DNA Testing Handbook - Advantage Medical Products

Transcription

Setting the Standard for Quality DNA Identification
DNA Reference Guide
A Collection of Technical Bulletins
Table of Contents
40-072.4
Topic
How Laboratory Accreditation Protects You
Legal Paternity Testing
Sample Collection and Chain of Custody
DNA Paternity Laws
High Resolution Paternity Testing
Probabilities in Paternity and Relationship Testing
Prenatal Paternity Testing
Understanding DNA Paternity Results
Understanding DNA Siblingship Results
Twins
Y Chromosome – A Tool for Paternity
Relationship Testing of Deceased Individuals
DNA from Exhumed Bodies
Forensic Paternity
DNA Relationship Testing in New York
Laboratory Procedures for Relationship Testing
DNA Mutations Explained
NADCM
ChromoCONNECTTM
Immigration
MiniFiler for Inhibited or Degraded Samples
NGM PCR Amplification Kit
Infidelity Testing
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How Laboratory Accreditation Protects
You
Technical Bulletin 40-053.1
The results of a paternity test, whether for legal purposes or
peace of mind, can result is life changing events for the
tested individuals. Because of the significance these results
play in individual’s lives, it is extremely important to ensure
that strict procedures are followed to ensure accurate
results. The cost of incorrect results can cause irreparable
harm to families and exposure to legal liability.
Partnering with an AABB accredited laboratory helps ensure
that the results generated are in compliance with the
highest possible peer reviewed standards.
Major Requirements of AABB Accreditation
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Quality and Management System
Regular internal and external audits
Regular proficiency testing of DNA analysts
Standard Operating Procedures
Stringent Qualifications for Staff
Use of Validated Methods
Duplicate Testing of Exclusions
Continuing Education for Staff
Regular Calibration of Laboratory Equipment
Record retention requirements
Confidentiality Polices
The Chromosomal Quality System
A core philosophy at Chromosomal Labs ● Bode Technology is “Quality Built-In”. This means that every step of the
process, from sample collection to results reporting, is critically designed, evaluated and monitored. This translates
into the highest quality possible.
The Quality System at Chromosomal Labs ● Bode Technology is modeled after ISO 17025:2005, General
Requirements for the Competence of Testing and Calibration Laboratories, the international benchmark for
approving the competence of testing and calibration laboratories. The Chromosomal Labs ● Bode Technology
quality program has been engineered to meet the diverse requirements of several accreditation bodies and
standards, including AABB, ASCLD and DAB. The laboratory complies with applicable industry guidelines, including
the Quality Assurance Standards for Forensic DNA Testing Laboratories and Convicted Offender DNA Databasing
Laboratories issued by the FBI Director.
Accreditation – The Cost and Value of Quality
External laboratory accreditation and proficiency testing programs are expensive to administer and require a firm
commitment to quality and excellence. Because of the life changing impact that a paternity test report can have, the
selection criteria of an analytical laboratory should have a strong emphasis on accreditation and quality.
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Email: [email protected]
Website: advantagemedicalproducts.com
(520) 750-7555 - Tucson, AZ
Toll Free: 1800-211-4433 - Outside Tucson, AZ
Legal paternity tests can be used in a court of law and are often
performed when one or both of the parents are seeking custody, child
support or social security payments. A legal test should be performed
in any circumstance in which the test participant(s) think they may want
to take the results to court or use them in a legal proceeding.
For a legal test, specific collection, consent, documentation of samples,
identification records and chain of custody must be maintained as they
relate to the case.
Sample Collection
Collection materials shall only be sent directly to collectors and/or witnesses. Collection materials shall not be in the
possession of any of the tested parties either before or after collection. Samples must be collected by an individual
with no interest in the outcome of the test. Tested parties can not self-collect. In addition, the samples must be
analyzed by a laboratory accredited by the AABB. Individuals qualified to collect samples include third party
administrators, physicians, child support workers, attorneys, ministers, or a person/company hired by the laboratory.
Consent
For a legal case, it is always imperative to obtain and document consent of the tested individuals.
Documentation of Samples
Each sample envelope shall be labeled with the name of the individual being sampled and the stated relationship (i.e.,
mother, child, and father). The envelope shall also document the date the sample was collected.
After collecting the samples, the sample collector and the person being sampled (or the individual with legal authority)
shall sign the sample envelope verifying that the samples inside belong to the person whose name is on the sample
envelope.
Identification Records
Both personal information and legible photo-identification must be collected and submitted to the laboratory in order to
process a legal case. The following personal information is required for each individual being tested:
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Printed name (must be legible)
Alleged relationship (i.e. mother, child, father)
Date of birth
Race/ethnicity (with the exception of the child)
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Location of sample collection
Date of sample collection
Printed name and signature of the person collecting the sample or witnessing the collection
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A history of any blood/bone marrow transfusion/transplant.
Photo-Identification
It is required that a legible photograph AND/OR copy of a government-issued photo ID be supplied at the time of
collection.
All tested individuals may appear in a single photograph together or they can be photographed individually. The
photos are used to positively ID the person that was sampled for the test, so that the individual(s) being sampled
cannot send another person in their place. To help prevent fraud, it is always best to photograph the test participants
at the time of collection. Individuals may not always look like their driver’s license picture, and therefore a driver’s
license can easily be given to a friend to use for a sample collection.
In order for a test to be legal, it is imperative that either a copy of the government photo ID or photograph be
submitted with the samples. It is also good practice to have the tested individual(s) and collector sign and date the
photographs, but it is not required.
If a government issued photo-ID or photograph cannot be obtained, either because the sample is from a deceased
individual, a prenatal sample, an incarcerated individual or a sample provided by law enforcement officials, a signed
affidavit attesting to the identity of the individual sampled, may be supplied by the individual who performed or
witnessed the sample collection, such as a doctor, coroner, corrections officer, etc. Some laboratories may have a
special form that can be filled out for these particular situations.
Chain of Custody
Chain of custody is documentation of where samples have been and who has handled them. Chain of custody begins
as soon as a sample is collected. The person collecting the sample has ownership of the samples until they transfer
them to the next person, usually a shipping carrier, such as FedEx, DHL, UPS, etc.
Do not allow test participants to package or transport samples. After samples are collected they should be placed into
tamper evident envelopes, sealed and given to a carrier for shipment to the laboratory. This action can be
documented on the chain of custody form by signing the name, date and time that samples were transferred from one
individual to the next.
When the laboratory receives the samples, they will continue the chain of custody by documenting how they received
the samples, who received them and at what time.
Chromosomal Labs ● Bode Technology is accredited by AABB for relationship testing. In accordance with AABB
standards, all exclusions are run in duplicate. In addition, every report is reviewed by a Ph.D. scientist. Chromosomal
Labs ● Bode Technology prepares a court admissible package for legal tests which includes a legal
paternity/relationship report, chain of custody form and photo-documents of the tests participants. Should the need
arise, Chromosomal Labs ● Bode Technology also offers expert witness testimony for a fee.
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WHAT IS CONSIDERED A LEGIBLE PHOTOGRAPH?
For legal cases, AABB requires LEGIBLE photographs, which can be either a government-issued photo
ID or a photograph that is suitable for positive identification.
To help clear up any confusion in regards to what Chromosomal Labs ● Bode Technology believes
constitutes a legible photo, examples of acceptable and unacceptable photos are shown below.
ACCEPTABLE
UNACCEPTABLE
UNACCEPTABLE
Photos do not have to be printed on photo paper, nor do they need to be in color, they simply need to be legible.
If a photo is not legible (i.e. the photo is too light or too dark) and the individuals sampled cannot be positively
identified from the photograph, a new photograph will need to be submitted. Legal reports cannot be released without
a legible photograph. Please double check your photograph(s) before your customers leave, so that you have an
opportunity to take another photograph, should the first one be illegible.
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Sample Collection and Chain of Custody
The value and defensibility of a laboratory DNA relationship
report is dependent in part on sample collection
procedures/techniques and on an unbroken chain of
custody. Proper sample collection and chain of custody
procedures are essential to ensure the accuracy of the
analysis as well as to ensure that proper documentation has
taken place for potential use in legal proceedings. Failure to
correctly follow either procedure may nullify or disqualify not
only the accuracy of the analyses, but also their use as legal
evidence.
The type of paternity test you require will depend on the reason(s) you are having the test performed. If it is simply for peace of
mind, you can utilize the non-accredited private paternity test, however if it is regarding such areas as child support, child
custody, or immigration applications you will want to utilize an AABB accredited legal paternity test. The primary difference
between the two types of tests is the sample collection method which makes private paternity tests non-accredited. Private
paternity test samples can be self-collected without any witnesses, while legal paternity test samples are collected at an
approved collection site, where your identification will be verified, a photograph will be taken and your sample collection will be
witnessed. The purpose of these stringent procedures is to ensure that the participants are who they say they are and to make
sure that accurate samples have been taken to submit to the laboratory.
The following bullet points all describe required sample collection/chain of custody procedures for a legal paternity test.
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The laboratory will have all necessary policies, procedures, and processes for proper sample collection and
maintenance of identification documents.
Each person tested will provide informed consent before sample collection. In the case of minors, the individual with
legal authority will provide informed consent.
A competent person, with no interest in the test outcome, will perform the sample collection and document the process.
o This person is responsible to protect the sample from contamination, protect the safety of the individual being
sampled, and maintain sample integrity.
The sample collection professional must confirm:
o The person(s) being tested have provided appropriate identification and the relationship of the individuals is
accurately recorded.
o Informed consent has been obtained.
o The sample was collected from the proper individual.
o All sample labeling is accurate.
o The sample(s) are properly packaged in a tamper-evident manner.
Each sample requires a unique identification, the date of sample collection, and the sample collector’s initials.
The accuracy of the labeling will be documented by the sample provider or individual who gave legal consent for the
person being sampled.
A legible photo of each individual collected must be take and properly labeled and signed.
The samples must be transported to the laboratory in a tamper evident envelope or package.
o Individuals tested cannot package or transport the samples
The printed name, birth date, and alleged relationship of each individual tested must be recorded.
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DNA Paternity Laws
US Paternity Laws can trace their origins to 18th Century
England. Chief Justice of the King’s Bench, Lord William
Murray Mansfield, served from 1756 until his retirement at
age 80 in 1788. His decisions greatly influenced English
common law, which remains the basis of law in the United
States today. Sometimes referred to either as the Bastardy
Statute or British common law origin, the Lord Mansfield's
Rule, defines a child born into a marriage to be a product of
that marriage. The husband of the mother is legally bound
to support the child as though it were his own. While use of
the Mansfield Rule is falling on disfavor, many states still
use this guiding principle.
Establishing Paternity
Establishing paternity means being declared the legal father after the child is born. If the parents of a child were not
married when the mother became pregnant or when the child was born, the child does not have a legal father until
paternity is established.
Establishing paternity is important for both the child and the parents. A child is entitled to the sense of belonging
and identity that comes from knowing both parents. Establishing paternity often has an important emotional and
psychological benefit for the child. Also, until there is a determination that a biological father is the legal father, the
child has no right to receive financial support from the father, to inherit from the father or to obtain insurance,
veterans’, social security or other benefits through the father. Likewise, until paternity is legally established, the
biological father has no legal rights to the child, such as parenting time with the child. Additionally, he has no legal
right to participate in major decisions about the child, such as medical treatment, education or religious training.
Additionally, genetic diseases are inherited, thus knowing your biological ancestry can be useful in proactive care
and transplantation.
Paternity laws tend to vary from state to state. The Uniform Parentage Act of 2002 by the National Conference of
Uniform State Laws modernizes the law for determining the parents of children, and facilitates modern
methods of testing for parentage.
History of Paternity Testing
Before DNA testing became available, several blood testing methods were used to determine paternity. These tests,
based on different blood group systems (BGS), were difficult to perform and often produced inconclusive results.
Most courts now accept only DNA test results as evidence for paternity cases.
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DNA Paternity Laws
Evolution of Paternity Testing Methods
Time
Period
1920s
1930s
1970s
1980s
1990s
Method
Sample
ABO Blood Type
Rh, Kell and Duffy Blood Groups
HLA Typing
RFLP – DNA
PCR – DNA
Blood
Blood
Blood
Blood
Swab
Power of
Exclusion
30%
40%
80%
99.99%
99.99%
ABO Blood Typing
Certain proteins located on the surface of red bloods cells, called ABO antigens, determine the blood type of A, B,
AB or O. ABO antigens are inherited from the father and the mother. Depending on the blood types of the two
individuals it was sometimes possible to exclude an alleged father, but not prove paternity.
Blood typing, based on the ABO blood group system, is not an accurate method for determining paternity. It
eliminates (excludes) only 30% of the entire male population from being the possible father. It cannot be used to
prove paternity.
Rh, Kell and Duffy Blood Groups
This test is somewhat similar to the ABO typing system in that it measures inheritable protein groups in blood. This
method increased the power of exclusion to approximately 40%, but was not useful in proving paternity.
HLA Typing
HLA Typing measures Human Leukocyte Antigens (HLA), found on most cells in the body, except red bloods cells.
White bloods cells contain the greatest concentration of HLA and are the same markers associated with organs
transplant rejection.
This method increased the power of exclusion to 80% and in some cases it was possible to produce a probability of
paternity of up to 90%. However, HLA testing could not differentiate between related alleged fathers. The large
amount of blood required for this test limited its use to infants older that 6 months of age, and made testing
uncomfortable for small children.
RFLP
RFLP, short for Restriction Fragment Length Polymorphism, is a DNA technique that relies on the genetic
uniqueness of all individuals in the world. No two persons have the same DNA sequence, except for identical twins.
DNA is extracted from blood and treated with specific enzymes to create DNA fragments. The size of the fragments
are measured and statistically analyzed. If too many fragments do not have a match, then the father is excluded.
This technique increased the power of exclusion to 99.99% and greater.
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DNA Paternity Laws
PCR
PCR, or Polymerase Chain Reaction, is similar to RFLP in that it uses DNA. PCR is an extremely powerful
technique for analyzing DNA on very small amounts of biological samples, from almost every part of the body. All
cells in the body have the same DNA, so the results are the same regardless of the type of biological sample taken.
PCR produces billions of copies of select portions of DNA from a small sample, such as a swab rubbed against a
patient’s inner cheek. The resulting DNA is analyzed on a Genetic Analyzer and statistically evaluated. This
technique has a power of exclusion of 99.99% or greater.
PCR has been extensively used for DNA testing, and large databases have been accumulated for accurate DNA
analysis. This large database enables paternity testing via PCR to have the highest power of exclusion.
Paternity Fraud
Paternity fraud, or the false identification of a man as the father of a child, is a serious problem in the US and
throughout the world. According to USA Today, a report from the American Association of Blood Banks says that
approximately 30% of the paternity tests performed annually in the United States result in exclusion of the alleged
individual as the biological father. Today, it is projected that the annual number of persons that will participate in
some type of paternity or extended relationship test will exceed 1 million.
The LA Chapter of the National Coalition of Free Men estimates that there are over 7 million children in the United
States that are unknowingly calling the wrong person, Dad.
Paternity Fraud or the false identification of a man as the father of a child, forces thousands of men each year to pay
for children fathered by other men. In many cases paternity is assigned by default without testing or appearance of
the accused. Recent legislative trends in some states are providing justice for the wrongfully accused.
Method for Determining Paternity
Almost universally, paternity testing is now performed using PCR. When selecting a paternity test, there are two
main factors to consider.
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Number of Genetic Markers Tested
Type of Paternity Test (non-accredited Private or AABB accredited Legal)
Genetic Markers
DNA results often decide the outcome of the case. It is important to understand the bottom line results on the
paternity test report. All paternity test results and conclusions are founded in statistics and probabilities. The
greater the number of genetic markers examined the greater the strength of the genetic evidence and hence the
final result. Industry wide, paternity tests can be generated using as few as four markers, to as many as 16
markers, with the latest high resolution technology. The 16 marker technology can achieve a probability of identity
of 1 in 40 quintillion, or 1 in 40,000,000,000,000,000,000.
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DNA Paternity Laws
Private Paternity Test
This self administered procedure can be accomplished by testing only the alleged father and the child. However, it
is always recommended that the mother be included, if possible. Including the mother will always improve the
results. Collecting samples for DNA analysis is an easy and painless procedure. A sterile swab, very similar to a Qtip, is rubbed on the inside of the cheek from the child and the alleged father. The swabs are then placed into their
corresponding envelopes and sent back to the lab for analysis.
Private paternity tests are excellent for cost effective confidential determination of paternity for a number of
applications including peace of mind, curiosity, or validating a case prior to incurring the expense of court orders and
attorneys. Private paternity testing is not available in the State of New York. Private (non chain of custody) paternity
tests are not accredited by AABB at any laboratory.
Legal Paternity Test
A Legal Paternity test is very similar to a private paternity test. The primary difference is that the samples must be
collected by a competent person with no interest in the outcome. This means that spouses, children, family or
friends are excluded from collecting samples. People that may qualify include ministers, child support workers,
physicians, attorneys or a person hired by the laboratory. At the time of sample collection, identification will be
verified and a photograph will be taken and sample collection will be witnessed. Legal paternity tests are required
for consideration by the courts and are accredited by AABB.
Private versus Legal Paternity
Criteria
Private Paternity
Court Admissible
No
Confidential
Yes
Painless
Yes
Court Ordered
No
Requires 3rd Party Sample Collection
No
Convenient
Yes
Accurate
Yes
In-home testing
Yes
Accredited by AABB
No
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Legal Paternity
Yes
Yes
Yes
Yes
Yes
No
Yes
No
Yes
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DNA test results often decide the outcome of a paternity
case. All paternity test results and conclusions are founded
in statistics and probabilities. The greater the number of
genetic markers examined the greater the strength of the
genetic evidence and hence the final result. Industry wide,
paternity tests can be generated using as few as four (4)
markers, to as many as sixteen (16) markers, with the latest
high resolution technology. The 16 marker technology can
achieve a probability of identity of 1 in 40 quintillion, or 1 in
40,000,000,000,000,000,000.
DNA paternity test results are typically expressed by three
sets of numbers, the paternity index (PI), the combined
paternity index (CPI) and the probability of paternity (W).
Paternity Index (PI)
Paternity index or likelihood ratio is the statistic that summarizes all of the information provided by the genetic
testing and is defined as the probability that some event will occur under a particular set of assumptions divided by
the probability that the same event will occur under a different set of mutually exclusive assumptions. For example,
in the case of paternity, the primary assumption is that the results for an individual genetic marker support the
assumption that the tested man is the true biological father rather than an untested randomly selected unrelated
man.
Combined Paternity Index (CPI)
The combined paternity index is determined by multiplying the paternity index values for each genetic marker
tested. The value can range from 0 to infinity. Values less than 1 are indicative of non-paternity or non-kinship and
if less than 0.001, exclusion of paternity. A value of one is neutral and does not provide evidence either way.
Values greater than 1 suggests the tested man is the father. The greater the CPI value, the stronger the stronger
the genetic evidence. Most states accept a CPI of 100 or greater as the standard to establish paternity.
Probability of Paternity (W)
The probability of paternity is an expression of the strength of one’s belief in the hypothesis that the tested man is
the father, based on all evidence in the case, including nongenetic evidence.
.
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The Effect of the Number of Genetic Markers Examined and the Combined Paternity Index
No.
Genetic
Marker
Child
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
D8S1179
D21S11
D7S820
CSF1PO
D3S1358
TH01
D13S317
D16S539
D2S1338
D19S433
vWA
TPOX
D18S51
D5S818
FGA
AMEL
12 13
28 31.2
8
11
10 13
15 17
6
9.3
8
12
9
11
17
14 16.2
14 18
8
15 17
11 14
21 25
X
Alleged
Father
13
28
8
12
15
6
8
11
17
13
18
8
12
12
21
X
9
13
17
9
11
12
25
14
17
14
Y
Mother
Paternity
Index
(PI)
12
5.74
28 31.2
6.4
8
11
1.1
10
5.89
15 17
1.65
6
9.3
2.67
11 12
2.11
9
12
1.62
16 17
6.59
13 16.2
2.11
14
5.6
8
1.87
14 15
4.67
11 12
22
25
7.72
X
1
Combined
Paternity
Index (CPI)
6
37
40
238
393
1,049
2,212
3,584
23,620
49,838
279,093
521,903
2,437,288
53,620,343
413,949,047
413,949,047
Relationship between CPI, Probability of Paternity and Population Frequencies
Combined Paternity
Index (CPI)
20
35
50
100
1,000
10,000
100,000
1,000,000
10,000,000
100,000,000
Probability of
Paternity (W)
95%
97%
98%
99%
99.9%
99.99%
99.999%
99.9999%
99.99999%
99.999999%
Number of Individuals in the
Population that Could Contribute the
Same Genetic Profile
1 in 20
1 in 35
1 in 50
1 in 100
1 in 1,000
1 in 10,000
1 in 100,000
1 in 1,000,000
1 in 10,000,000
1 in 100,000,000
11
5%
2.9%
2%
1%
0.1%
0.01%
0.001%
0.0001%
0.00001%
0.000001%
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Probabilities in Paternity and
Relationship Testing
DNA test results for paternity and kinship studies are
typically expressed by three sets of numbers.
Paternity results use the paternity index (PI), the
combined paternity index (CPI) and the probability of
paternity (W). Kinship results are expressed similarly;
however the term likelihood ratio, or LR, is utilized in
lieu of paternity index.
Paternity Index (PI) or Likelihood Ratio (LR)
Paternity index or likelihood ratio is the statistic that summarizes all of the information provided by the genetic
testing and is defined as the probability that some event will occur under a particular set of assumptions divided by
the probability that the same event will occur under a different set of mutually exclusive assumptions. For example,
in the case of paternity, the primary assumption is that the results for an individual genetic marker support the
assumption that the tested man is the true biological father rather than an untested randomly selected unrelated
man.
Combined Paternity Index (CPI) or Cumulative Likelihood Ratio (CLR)
The combined paternity index or cumulative likelihood ratio is determined by multiplying the paternity or likelihood
ratio index values for each genetic marker tested. The value can range from 0 to infinity. Values less than 1 are
indicative of non-paternity or non-kinship and if less than 0.001, exclusion of paternity or kinship. A value of one is
neutral and does not provide evidence either way. Values greater than 1 suggests the tested man is the father or is
evidence for kinship. The greater the CPI value, the stronger the stronger the genetic evidence. Most states accept
a CPI of 100 or greater as the standard to establish paternity.
Probability of Paternity (W)
The probability of paternity is an expression of the strength of one’s belief in the hypothesis that the tested man is
the father, based on all evidence in the case, including non-genetic evidence.
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The paternity of an unborn child can be determined
utilizing the same laboratory methods used in
conventional paternity testing. Prenatal DNA from
an unborn child is collected and compared to DNA
from the mother and the alleged father, whose
DNA is collected using conventional buccal swab
techniques. The prenatal DNA can be collected by
a physician utilizing one of two techniques
amniocentesis or chorionic villus sampling (CVS).
The type of sample collected will depend on the
stage of pregnancy. CVS is typically performed
between the 10th and 12th weeks of pregnancy,
while amniotic samples are collected between 12
and 21 weeks.
Prenatal Sample Collection
Amniocentesis is an invasive procedure that involves inserting a needle into the mother’s abdomen to
withdraw a small amount of amniotic fluid from the amniotic sac which surrounds the developing fetus.
This procedure can lead to damage or loss of the baby in up to 1% of cases. As a result, it is
recommended that patients wait until the baby is born, unless they are already undergoing this procedure
for medical reasons.
In general, a 10 ml sample of amniotic fluid is requested. Samples can be collected in sterile nonpreserved vials. Samples collected prior to 18 weeks gestation may contain insufficient fetal DNA for
analysis. Samples should be shipped frozen on blue ice or dry ice to the laboratory via overnight courier.
Chorionic villus sampling involves inserting a catheter through the cervix and removing portions of the
chorionic villi. Chorionic villi are microscopic finger-like projections that emerge from the chorionic
membrane and eventually form the placenta. The cells in the chorionic villi are of fetal origin, and thus
have the same genetic composition as the baby. This procedure can lead to damage or loss of the baby.
As a result, it is recommended that patients wait until the baby is born, unless they are already
undergoing this procedure for medical reasons. Samples should be shipped frozen on blue ice or dry ice
to the laboratory via overnight courier.
Newborn Sample Collection
Unlike older methods, a DNA paternity test can be conducted on individuals at any age, including
newborns. Samples collected at the time of delivery are typically done using umbilical cord blood
obtained at the time of delivery. This procedure involves the collection of the child's blood from the
umbilical cord at birth. Samples can be collected in lavender top tubes containing EDTA. If a sample is
collected after the birthing process, buccal swabs can be used.
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Cell Free Fetal DNA in Maternal Blood
There are a handful of companies that are offering a non-invasive prenatal paternity test. This test is
based on a 2005 article that appeared in the journal Science. The article presented research that
showed that cell free fetal DNA is present in maternal blood and can be recovered for DNA testing.
Shortly after this article appeared, Chromosomal Labs ● Bode Technology evaluated the science and the
labs that were offering it at the time and came to the conclusions there were a number of errors in the
results from those companies offering it. While the methods are suitable for the research arena, they
have not been properly validated as a diagnostic. A prenatal geneticist at Tufts University School of
Medicine stated that fetal DNA can remain in the mother’s blood stream for up to 20 years.
Chromosomal Labs ● Bode Technology does not offer this test, nor does it recommend laboratories that
offer it.
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Understanding DNA Paternity
Results
During a paternity test, a DNA profile is generated from each sample and will ultimately help prove or disprove the
paternity of a child.
The laboratory report will generally have one of three possible conclusions, which are determined from the DNA
results and a series of complex statistical calculations. Following is a statement of the most common conclusions
along with a simple explanation of its meaning.
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The results indicate that the alleged father is not the biological father of the child. This conclusion is based
on the absence of two or more obligate paternal alleles in the alleged father, as indicated by a Paternity
Index of 0.
This means that the tested man is not the father.
The results indicate that the alleged father can not be excluded as the biological father of the child. The
reported probability of paternity, as compared to an unrelated, untested man of the same race, is calculated
assuming a prior probability of 0.5.
The only way to be 100% certain that the alleged father is the true father would be to test every man on
earth. This of course is not possible. Because the probability is not 100% you cannot say in your
conclusion that the alleged father is the biological father, as this indicates 100% certainty. To determine
what “can not be excluded” means, think of it this way. If you tested 10 men as possible fathers, one would
be the father and 9 would not. If you lined them up in a room and went down the line, starting with father
#1, you would say to him “you are not the dad, you are excluded”, then to father #2, “you are not the dad,
you are excluded”, and down the line you go, until you get to dad #10 and you say “you CAN NOT be
excluded. So you cannot say he is the dad 100%, but you can say, that he can not be excluded as the
father.
The statistical results derived from the genetic tests fall within the inconclusive range for paternity.
Inclusion of the mother will likely be sufficient to yield a conclusive result.
This means that additional testing is necessary to determine if the tested man is the father or not the father.
This result occasionally occurs when the mother is not included in the analysis.
Paternity Index
Paternity index is a likelihood ratio between the chances that the alleged father may pass the paternal gene,
compared to the chance that a random man may pass the paternal gene to the child. If the paternity index is zero,
it is because the father does not have any matching alleles with the child at that particular marker. If the paternity
index is greater than zero it means the father does have a matching allele with the child.
Combined Paternity Index (CPI)
The combined paternity index is the product of all of the individual paternity index values multiplied together and is
a measure of the strength of the genetic evidence. The CPI can range from 0 to infinity. When the combined
paternity index is 0, it means the alleged father is not the dad. When the combined paternity index is greater than
100 it means the alleged father is the dad, however it cannot be stated simply as “you are the father”. Because the
probability of paternity can never be 100% (you will often `see 99.999%), you cannot say “you are the father”
because that implies 100% certainty. Therefore, the results are reported as “the alleged father can not be excluded
as the biological father of the child”. When the combined paternity index is greater than 0, but less than 100, the
results are inconclusive. Inconclusive results are most often the cause of doing a paternity test without the mother.
In most instances, inclusion of the mother’s genetic profile into the statistical calculations results in a conclusive
result.
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Understanding DNA Paternity
Results
The Science of Inheritance
The colored images below illustrate how DNA is passed down from the mother and father to the child. As you can
see, the mother passes a red chromosome to the child, and the father passes a blue.
Mother
Allele A =14
Allele B =15
Child
Allele A =14
Allele B =16
Father
Allele A =16
Allele B =16
Example Results for a Paternity Inclusion
Child
Alleged Father
Mother
Genetic
Marker
Allele A
Allele B
Allele A
Allele B
Allele A
D8S1179
D21S11
D7S820
CSF1PO
D3S1358
TH01
D13S317
D16S539
D2S1338
D19S433
vWA
TPOX
D18S51
AMEL
D5S818
14
30
10
11
15
7
11
10
24
14
14
8
14
X
11
15
16
30.2
13
11
17
13
14
30
10
10
16
7
11
10
22
14
14
6
14
X
11
12
12
24
16.2
17
11
15
Y
13
16
30.2
10
10
16
6
12
12
18
14.2
15
6
13
X
11
14
22
16.2
17
8
15
Y
13
1.81
38.5
23.4
3.66
3.24
1.13
2.06
1.66
3.56
23.9
2.69
5.66
2.12
1
1.96
FGA
24
28
24
26
19
26
12.9
12
12
17
9
13
12
25
18
11
15
Combined Paternity Index
Probability of Paternity
Allele B
31
13
7
Paternity
Index
5,190,000,000
>99.999%
Discussion
If the father’s DNA profile does not contain a matching allele with the child the paternity index would be zero. When
two or more genetic markers yield a paternity index of zero, the alleged father is excluded as the biological father of
the child. In this case the alleged father has a matching allele at every marker, so he can not be excluded.
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DNA Siblingship Testing
A DNA siblingship test determines the likelihood that two
individuals are related siblings, either as full siblings or as half
siblings.
Applications
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Two individuals are known to share one biological
parent, and want to determine if they share the second
parent
Two individuals want to determine if they share one
biological parent
An alleged father is unavailable for a paternity test
Adoption reunion
Siblings who were separated at birth
Immigration DNA Testing
The Science of Inheritance
The colored chromosomes below illustrate how DNA is inherited from the mother and father. The mother
passes a pink chromosome to child 1 and a green chromosome to child 2. The father passes a blue
chromosome to both children.
In a DNA siblingship test, many different chromosomes are analyzed. If two individuals are related as full
siblings, approximately 50% of the DNA results should be the same. If two individuals are related as half
siblings, approximately 25% of their DNA should be the same. During a DNA siblingship test, at least 16
different DNA markers are evaluated. The number of shared markers and the pattern in which they are shared
are analyzed, and a relationship index is calculated.
Mother
Allele A =14
Allele B =15
Child 1
Allele A =14
Child 2
Allele A =16
Allele B =15
17
Father
Allele B =16
Allele A =16
Allele B =16
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Siblingship DNA Test Results

Likelihood Ratio
The likelihood ratio is a measure of the likelihood of a sibling relationship between two tested individuals.
Because of the complexity of a siblingship test, the results are not as definitive as those of typical
paternity tests.
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Combined Likelihood Ratio
The combined likelihood ratio is the product of the individual relationship index values and is a measure
of the strength of the genetic evidence.
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Result Interpretation
In sharp contrast to DNA paternity testing, there are no obligatory genes in siblingship tests. As a
consequence, the results of siblingship testing are reported as the likelihood in support of or against an
alleged relationship. While interpretation of the strength of the statistical value can be variable, and
should ultimately be considered in context with all case circumstances, the table below summarizes
published interpretative criteria for use as a guide.
<1
1
1 to 10
10 to 100
100 to 1,000
1,000 and greater
Support of Siblingship Hypothesis
Genetic evidence does not support hypothesis
Neutral
Limited Support
Moderate Support
Strong Support
Very Strong Support
Increasing the Discrimination Power of Siblingship Tests
In cases where the standard 16 genetic marker panel does not yield an adequate result, extended testing can be
performed to obtain a statistically desirable result. Extended testing options can include testing the mother or
mothers if they are known, adding more autosomal genetic loci, or Y chromosome testing, if both children are
male. Please contact a DNA Case Manager to review possible extended testing options.
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Twins
Approximately one in every fifty Americans is a twin, two
thirds of which are fraternal and one third identical.
Triplets occur once in every 8000 deliveries.
Twin classification done at delivery often results in
misidentification. For example, approximately 25-33% of
identical twins have two sets of chorionic membranes.
Unless a DNA or other blood test was performed, it is
difficult to accurately determine if twins are fraternal or
identical.
Identical twins are formed when one egg is fertilized by a single sperm cell and then divides within the first few
days of pregnancy. If division of the egg occurs after 8 days, this can result in conjoined twins, also known as
Siamese twins. Twins produced by this process are referred to as identical since their DNA sequence is identical.
While the twins are identical genetically, this often does not apply to the actual physical or behavioral traits.
Generally their traits are similar, however they can differ. The occurrence and distribution of identical twins in the
population appears randomly and does not matter when, where, or to whom they were born.
Fraternal twins are formed when two eggs are independently fertilized by two sperm cells. Fraternal twins do not
have identical DNA but will have half of their DNA in common, just like siblings of the same parents that are born
at different times. Fraternal twins can either be of the same sex or different sex.
The occurrence of fraternal twins varies substantially in frequency around the world. They are common in the
pregnancies of older women, especially common in Africans, and relatively rare among those of Asian heritage.
Twins of the World
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Forensics
DNA crime scene evidence from an identical
twin can hinder prosecution because two
people have identical genetic markers.
Identical: 33%
Fraternal: 67%
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Y Chromosome – A Tool for Paternity
Testing
Conventional paternity testing utilizes diploid
genetic markers and takes advantage of the fact
that each individual is a mosaic of their mother and
father, resulting from a blending of the DNA it
receives from each. This approach is extremely
accurate when the father is available for testing.
In the absence of the alleged father, family
reconstruction is historically conducted through
siblingship, avuncular or grand paternity, which can
at times yield inconclusive results. In certain
scenarios, Y chromosome testing can be an
extremely powerful technique for paternity and
other relationship testing.
Y Chromosome
The Y chromosome is male specific and passes through generations unchanged from father to son.
In
circumstances where an alleged father is unavailable for testing or deceased, a male child can be tested against
any number of his male relatives including, but not limited to, other known male children, brothers and half brothers
with the same father, uncles, grandfather, and grandfather's brothers.
Limitations
Since the Y chromosome is only found in males, this type of testing is not useful for female off-spring. Similarly this
test is not iron clad for paternity in circumstances where there are two related alleged fathers.
Thomas Jefferson DNA Study
This technique was recently used to determine if Thomas Jefferson fathered Eston Hemings. The study compared
nineteen (19) genetic markers on the Y chromosome from five Jefferson descendants and one male line
descendent of Eston Hemings, Sally Hemings’ last son, born in 1808. Analysis of the Jefferson and Hemings lines
revealed an identical genetic match of a distinct genetic profile that is conserved within the Jefferson family.
The Thomas Jefferson Foundation, who owns and operates Monticello, commissioned a research committee to
evaluate the DNA study. The committee concluded there was a high probability that Thomas Jefferson fathered
Eston Hemings, and that he most likely was the father of all six of Sally Hemings’ children appearing in Jefferson's
estate records.
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Relationship Testing of
Deceased Individuals
In death, unresolved questions of biological relationships can impact surviving family members at a
number of different levels, including custody, inheritance, social security and public assistance.
DNA Testing
Recovery of DNA from remains is often highly successful, depending on the quality of preservation.
Usually a number of different sample types are collected so that in the event that one does not yield
suitable DNA the likelihood of successful DNA recovery from at least one sample will typically remain high.
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Buccal Swab - it is often useful to moisten the swab with sterile distilled water prior to swabbing an
embalmed person
Fingernail clippings from one hand
Blood or FTA blood card
Autopsy Tissue Section (1 cm)
Samples should be placed in a suitable container and shipped over night to the laboratory. If fresh tissue
is shipped, samples should be frozen and shipped on dry ice.
Legal Relationship Testing
Legal relationship testing involving deceased individuals has the same basic requirements as any other
legal test. However, consent requirements will be different and sample identification will come from the
facility that has current custody of the samples. To adjust for this, Chromosomal Labs ● Bode Technology
will provide you with the necessary forms to use. The third party administrator, next of kin or person with
legal authority* needs to contact the facility that has the samples or body of the deceased to make
arrangements.
1. The samples of the deceased individual must adhere to a strict chain of custody. The facility with
the samples will fill out the Chain of Custody/ Affidavit of Identity form. It is best to have the
pathologist, coroner or funeral home send the samples directly to Chromosomal Labs • Bode
Technology. It is not necessary for the third party administrator to take custody of the samples at
any time. The samples are never to be in the custody of the next of kin, or any other person
involved in testing.
2. The next of kin, or person with legal authority*, fills out the Third Party Client Authorization
form. This form gives Chromosomal Labs ● Bode Technology and its affiliates authorization to use
the samples of the deceased in a relationship test. This form must be submitted to Chromosomal
Labs ● Bode Technology prior to testing. Alternatively a court order, that clearly states that the
samples of the decedent be tested for paternity can be used.
*Legal Authority includes, Next of Kin, Power of Attorney, Executor of Estate, etc. For more information on who has legal authority
over the deceased and their samples consult your State Laws.
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Decomposition of a human body begins at the
moment of death and is initiated by two mechanisms,
autolysis, the breaking down of tissues by the body's
own internal chemicals and enzymes, and
putrefaction, and the breakdown of tissues by
bacteria. Secondary decomposition is accomplished
by scavengers, such as insects, provided the body is
accessible to them. The most common insects that are
typically involved in this process include the fleshflies
and blowflies.
The overall rate at which a body decomposes can vary greatly and is strongly influenced by a number of
factors. Some of the more important factors that affect the decomposition rate include:
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Temperature
Oxygen
Insects
Burial depth
Trauma
Humidity
Rainfall
Body size
Embalming
Clothing
An unembalmed adult body that is buried six feet deep in normal soil, without a coffin, takes approximately ten
to twelve years to decompose fully to a skeleton, given a temperate climate. If the body is immersed in water,
skeletonization occurs in approximately three years. If it is exposed to air, decomposition can occur within one
year. Bodies that are exposed to cool, damp soil may develop a waxy substance called adipocere, caused by
the action of soil chemicals on the body's proteins and fats. The formation of adipocere slows the rate of
decomposition by inhibiting the bacteria that cause putrefaction.
Embalming
Since the 1920s, it has been standard practice for American morticians to embalm corpses, to delay
decomposition. Embalming chemicals will repel most insects, and slow the process of bacterial putrefaction, but
will not preserve a corpse indefinitely. In dry environments, an embalmed body may become mummified.
Cremation
The process of cremation involves incinerating the body at a temperature of 1,400 to 2,100°F for approximately
two hours. During this process, the majority of the body is combusted and discharged through an exhaust
system. The remains after cremation are bone fragments, representing about five percent of the body's
original mass. Little forensic work has been done on cremated specimens; however, most experts believe it is
unlikely that DNA would survive the cremation process. While a Japanese team has reported obtaining a DNA
profile from cremated remains, the validity of their results remains controversial.
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Exhumation
Attitudes toward exhumation vary wildly based on religious or cultural beliefs. Some Native American tribes
believe that the body's spirit lives on and that to disturb a corpse is to disturb the spirit's life. Similarly, people of
both Jewish and Muslim faith are averse to disinterment. Christians tend to have a more liberal exhumation
policy. Both the Catholic and Protestant churches take the position that bodies shouldn't be disturbed, if
possible. Saints have frequently been disinterred, upon canonization, so their remains can be dismembered
and turned into relics.
In modern society, exhumations are relatively common to obtain DNA to settle paternity suits and for criminal
investigations.
U.S. laws governing exhumation are vague and disparate, varying from state to state. Most state regulations
were derived from English common law, which prohibits the theft of items from a grave, but does not address
theft or removal of bodies themselves. Historically, this was a matter left to the church. Only in the mid-19th
century, when snatching corpses for medical experiments became endemic, did states pass laws prohibiting
cadaver theft. Generally, people can apply to their state's attorney general for permission to exhume family
members for any reason. Family disagreement on exhumation typically must be resolved in the courts.
DNA Samples
Recovery of DNA from exhumed remains is reasonably successful, however, not guaranteed. Generally a
number of different sample types are collected, in the event that one does not yield suitable DNA.
Arrangements for sample collection can often be made through a funeral home or whoever exhumes the body.
Recommended sample types include the following:
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Fingernails
Tooth
Femur
Foot phalanx, patella, metatarsal
Tissue Section (1 cm) - Aortic tissue should be collected, if available.
Samples should be placed in a suitable container and shipped over night to the laboratory. If fresh tissue is
shipped, samples should be frozen and shipped on dry ice.
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Forensic Paternity
With conventional paternity testing it is customary to collect a voluntary or
court ordered buccal sample from an alleged father or individual for DNA
analysis. In certain circumstances it may be either difficult to confront an
individual or they might be deceased or otherwise not available to give a
DNA sample. In these circumstances, abandoned evidence can often be
useful in answering relationship questions. While the results will likely
not be court admissible, unless collected by a third party investigator,
they can provide valuable information on family relationships.
Sources of DNA Evidence
DNA is a component of virtually every cell in the human body and is constantly shed from a variety of sources,
including skin cells, saliva and hair. Forensically valuable DNA can be found on evidence that is decades old.
Several factors can affect DNA stability, such as sunlight, moisture, bacteria, and mold. Consequently, not all DNA
evidence will result in a usable DNA profile. Common sources of forensic DNA evidence include:
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Band Aids, feminine products, diabetic glucose
sticks
Blanket, pillow, bed sheet
Bone
Bottle, can, or glass
Dental floss
Dentures
Dirty laundry
Electric Razor Clippings
Eyeglasses
Facial tissue, cotton swab
Fingernail clipping
Gum
Hair w/ roots or follicles
Hat
Post mortem tissue
Stamp or envelope (lickable)
Teeth
Toothbrush
Toothpick
Used cigarette
Evidence Collection and Contamination Prevention
Because extremely small samples of DNA can be used as evidence, greater attention to contamination issues is
necessary when identifying, collecting, and preserving DNA evidence.
To avoid contamination of evidence that may contain DNA, always take the following precautions:



Wear gloves
Avoid touching the area where you believe DNA may exist
Avoid talking, sneezing, and coughing over evidence
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Forensic Paternity
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

Avoid touching your face, nose, and mouth when collecting and packaging evidence
Air-dry evidence thoroughly before packaging
Put evidence into new paper bags or envelopes, not into plastic bags
Transportation and storage
When storing and transporting evidence that may contain DNA, it is important to keep the evidence dry and at
room temperature. Once the evidence has been secured in paper bags or envelopes, it should be sealed,
labeled, and transported in a way that ensures proper identification of where it was found and proper chain of
custody. Never place evidence that may contain DNA in plastic bags because plastic bags will retain damaging
moisture.
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DNA Relationship Testing of Samples
Collected in the State of New York
State of New York Public Health Law
The New York State Public Health Law has strict
guidelines on DNA paternity and relationship testing.
Chromosomal Labs ● Bode Technology has a Clinical
Laboratory Permit, PFI 8237, from the New York State
Department of Health and can accept samples
collected in the State of New York for paternity and
relationship testing.
Paternity Testing
New York State regulations require that laboratories
test only under the authorization of a qualified person,
and to release results of laboratory tests only to
persons authorized by law to use such findings in their
professional practice or official duties, and not to the
case subject without written authorization from persons
qualified to order laboratory testing.
People Qualified to Order DNA Paternity Tests in the State of New York1
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New York judges ordering testing under the Family Court Act
New York Attorneys
Foreign Government (Consulate or embassy)
New York Licensed physicians
Physician’s assistants provided such examination is authorized by the supervising physician. If the request
is oral, the physician or other authorized person shall submit a written request to the laboratory within 48
hours. If the laboratory does not receive the written request within that time, it shall be noted in the record
of daily accession
Chain of Custody2
All paternity testing performed on samples collected in the State of New York shall meet standards for court
admissibility. New York State’s Public Health Law does not allow direct to consumer testing or private
testing.
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DNA Relationship Testing of Samples
Collected in the State of New York
Reporting of DNA Results3
According to Section 58-1.8
1. Results of tests are to be reported only to physicians or other authorized persons. No person shall report
the results of any test except to a physician, his agent, or other authorized person.
2. Reports shall not be issued to patients, except with written consent of the physician or other authorized
person.
Payment for DNA Testing4
Payment for testing can come from:
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
The tested individual or legal relative
A hospital
Governmental agencies on behalf of the tested individual
Attorney ordering and authorizing the test
Payment can not come from a third party administrator or physician
Disclaimer
The information contained on this page has been condensed from a number of official New York documents as a
courtesy and is believed to be accurate.
References
1. Persons authorized to order tests and receive directly the results of clinical laboratory testing of specimens
accepted from New York State, 6/22/06.
2. New York State Department of Health Clinical Laboratory Standards of Practice, Parentage/Identity Testing,
Standard PIT16.
3. PART 58-1 OF 10 NYCRRCLINICAL LABORATORIES, section 58-1.8.
4. New York State Public Health Law Title VI Laboratory Business Practices, Section 586.
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Laboratory Procedures for Relationship
Testing
Laboratory analyses are performed in accordance with validated standard operating procedures.
Samples are processed by one or more of the following techniques, depending on the nature of the
evidence and the analysis requested.
DNA Extraction and Purification: DNA extraction is performed by one of the following techniques:
EpiCentre QuickExtract, Helixtract, PrepFiler, Promega DNA IQ System, or proteinase K/phenolchloroform, depending on the nature of the sample.
DNA Analysis: Routine DNA amplification and fragment analysis is performed using the AmpFlSTR
Identifiler and/or Yfiler Kit and ABI PRISM® 3130xl Genetic Analyzer manufactured by Life Technologies.
Extended testing is performed using CS7 and PowerPlex 16 kits manufactured by Promega Corporation
and the NGM kit manufactured by Life Technologies. A summary of the actual STR loci analyzed are
presented with the laboratory results.
Statistical Analysis: Autosomal statistical calculations are performed using published allele frequencies
such as found in JFS, 2003, Vol. 44 (6), JFS 2001, Vol. 46, FSC 2001 Vol. 3, Promega.com and Applied
Biosystems. Y-STR profile frequencies were derived from www.appliedbiosystems.com/yfilerdatabase.
Analytical Platform
Paternity test results and conclusions are founded in statistics and probabilities. The greater the number
of genetic markers examined, the greater the strength of the genetic evidence and hence the greater the
reliability of the final result. Chromosomal Labs ● Bode Technology tests a minimum of 16 DNA markers
in routine relationship testing. The 16 marker technology can achieve a probability of identity of 1 in 40
quintillion, or 1 in 40,000,000,000,000,000,000.
Extended Testing
For the majority of paternity cases, the 16 genetic marker panel is more than adequate. However,
special circumstances can require extended testing to obtain a statistically desirable result. This typically
occurs when there is a single genetic mutation or the mother is not included in the test. Extended testing
options can include testing the mother, adding more autosomal genetic loci, or Y chromosome testing, if
the child is a male. During case review our doctors may determine that extended testing is necessary or
statistically useful, and will make recommendations for extended tests that are likely to be the most
productive.
Motherless Testing
While motherless paternity testing is a relatively common practice, it can present a number of problems,
ranging from inconclusive results, or in the case of incest or related alleged fathers, a false inclusion.
DNA testing of the mother, even if maternity is not disputed, improves the chance of obtaining conclusive
results and is a quality control check for both the scientific and legal community. In all relationship
testing, every effort should be made to test the mother when she is available.
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Laboratory Procedures for Relationship
Testing
Y-Chromosome Testing in Relationship Studies
The Y chromosome is male specific and passes through generations unchanged from father to son. In
circumstances where an alleged father is unavailable for testing or is deceased, a male child can be
tested against any number of his male relatives including, but not limited to, other known male children,
brothers and half brothers with the same father, uncles, a grandfather, and grandfather's brothers. Since
the Y chromosome is only found in males, this type of testing is not useful for female offspring. This test
is also not appropriate for paternity in circumstances where there are two related alleged fathers.
Genetic Inconsistencies (Mutations)
Genetic inconsistencies (mutations) are naturally occurring mismatches between a child and the alleged
parent. The frequencies of these naturally occurring mismatches are factored into the final combined
paternity index. Due to the nature of these calculations, the statistical value of the result is significantly
reduced; therefore some degree of extended testing is required to obtain a conclusive result.
Result Interpretation - Relationship Studies
The results of siblingship, avuncular and grand paternity testing are generally reported as the likelihood in
support of or against an alleged relationship. While interpretation of the strength of the statistical value
can be variable, and should ultimately be considered in context with all case circumstances, the table
below summarizes published interpretative criteria for use as a guide.
<1
1
1 to 10
10 to 100
100 to 1,000
1,000 and greater
Value of Evidence in Support of Hypothesis
Does not support hypothesis
Neutral
Limited Support
Moderate support
Strong Support
Very Strong Support
Quality Assurance
Chromosomal Labs ● Bode Technology is an AABB and ISO 17025 Accredited Relationship Testing
Facility. AABB is an international association involved in activities related to transfusion and cellular
therapies including transplantation medicine. Since its beginning in 1947, AABB continues to support the
highest standards of medical, technical and administrative performance, scientific investigation, clinical
application, standard setting, accreditation and education.
Chromosomal Labs ● Bode Technology is authorized to perform DNA Parentage/Identity Testing on
samples originating in the State of New York by the New York State Department of Health, PFI: 8237.
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DNA Mutations Explained
The human genome is very large, containing more than 3.2 billion bases, or letters. Despite its size, over
99.9% of the DNA in all unrelated people in the world is exactly the same. Consequently, the vast
differences observed in the human race are created from the differences in only 0.1% of DNA. These
differences are caused by mutations that have occurred gradually over the millennia.
As with any region of DNA, the STR genetic markers used in relationship and paternity testing are
subject to DNA mutations, also called genetic inconsistencies. They are naturally occurring mismatches
between a child and parent and can occur when a sperm cell is being made or when a cell divides. The
majority of STR mutations involve the gain or loss of a single unit. Thus, a maternal or paternal genetic
marker with a 14 would show as a 13 or 15 in the next generation following a mutational event. These
mutations can occur in either the mother or the father, but appear to be more frequent in the father. On
average, the genetic markers used in relationship testing mutate at an approximate rate of 0.13%.
The colored images below illustrate how a DNA mutation can be passed down from the father to the
child. As you can see, the mother passes one of her chromosomes to the child, and the father passes a
mutated chromosome that increased by one unit.
Mother
Allele A =10
Allele B =10
Child
Allele A =10
Allele B =15
Father
Allele A =14
Allele B =14
The frequencies of these naturally occurring mismatches are factored into the final combined paternity
index. Due to the nature of these calculations, the statistical value of the result is significantly reduced;
therefore some degree of extended testing is typically required to obtain a conclusive result. This usually
involves testing the mother if she was not included in the original test. Alternatively, additional autosomal
markers can be tested, or Y chromosome markers can be tested if the child is male and if the potential
alleged fathers are not paternally related.
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(520) 750-7555 - Tucson, AZ
National Association for
DNA Collection and
Management
Certified DNA Collection Technician

Industry Leading Certification

On-line Training Program

On-line Examinations

Free Technical Consultation

Standard Methods for the Collection of
Biological Specimens for Paternity and
Relationship Testing
The DNA Testing Market
The DNA relationship testing market has been growing steadily over the last twenty years, increasing on
average at a rate of 12% per year. Today, it is projected that the annual number of persons that will
participate in some type of paternity or extended relationship test will exceed 1 million. In sharp contrast,
it is estimated that less than 200,000 persons were tested in 1988.
The increased demand for testing has been fueled by greater public awareness of the power of DNA and
the accessibility of affordable DNA testing. The growth and popularity of the internet has also fueled
demand through increased direct to consumer marketing.
The accuracy of modern DNA testing methods is forcing a change to antiquated paternity laws across
the U.S. Many states are adopting laws that level the field for falsely accused men, allowing some
degree of financial relief from hardships created from paying support for another man’s child.
By all accounts, the outlook for the DNA paternity and relationship testing market is strong. As more
people take a proactive role in their lives, the DNA testing industry will further develop and mature and
ultimately require an increased demand for neutral third party administrators to provide DNA collections
and case management services.
Setting the Standard
A high growth industry tends to attract a dynamic range of players, each with a range of skill sets and
ethics. As the DNA testing industry evolves and the sophistication of the clientele increases, there will be
a need for firms that are offering DNA collection to differentiate themselves through quality benchmarks.
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(520) 750-7555 - Tucson, AZ
DNA Collection and
Management
The National Association for DNA Collection and Management (NADCM), a division of Chromosomal
Labs ● Bode Technology was created with that vision in mind.
Standardized Methods
NADCM’s Standard Methods for the Collection of Biological Specimens for Paternity and
Relationship Testing is a comprehensive sample collection resource that draws from a number of peer
reviewed publications representing industry best practice, including Standards for Relationship Testing
Laboratories, 8th edition, published by the American Association of Blood Banks (AABB), Forensic DNA
Typing, by John Butler, and regulations from New York State and U.S. Department of Homeland
Security.
State and federal requirements governing relationship testing laboratories in the United States are
variable. Except for the State of New York, which has its own formal relationship laboratory evaluation
program, the de facto industry standard is accreditation by the American Association of Blood Banks
(AABB) for relationship testing. This accreditation demonstrates conformance to the current edition of
Standards for Relationship Testing Laboratories. These standards have specific requirements for
sample collection that must be met for compliance. Consequently, third party administrators providing
sample collection services for relationship testing must be compliant with these standards, as well as
other industry best practice.
The Basics of DNA Paternity and Relationship Reporting is a technical resource designed to give
third party administrators (TPA) a general understanding of DNA and its uses in paternity and
relationship reporting. After obtaining this certification TPA’s will gain an understanding of how to
interpret paternity and relationship reports, mutations, Y Chromosome and Mitochondrial reports.
Following is an outline of the topics covered in this certification.
1.0
An Overview of DNA
1.1 Chromosomes
1.2 Polymerase Chain Reaction (PCR)
1.3 Electrophoresis
2.0
Paternity Testing and Statistics
2.1
2.2
2.3
2.4
2.5
2.6
Genetic Markers
Alleles
Paternity Index
Combined Paternity Index
Probability of Paternity
Conclusion Statements
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DNA Collection and
Management
2.7 Example Report – Paternity Trio – Inclusion
2.8 Mutation or Genetic Inconsistency
2.9 Example Report – Paternity Trio – Mutation
2.10 Example Report – Paternity Duo – Mutation
2.11 Example Report – Paternity Duo – Mutation #2
2.12 Example Report – Paternity Trio – Mutation #2
2.13 Exclusions
2.14 Example Report – Paternity Trio – Exclusion
3.0
Relationship Testing and Statistics
3.1 Grand Paternity
3.2. Example Report Grand Paternity – Mother, Child, 2 Grandparents
3.3 Example Report Grand Paternity – Child, 2 Grandparents
3.4 Avuncular
3.5 Avuncular (Motherless)
3.6 Siblingship
3.7 Siblingship – Full vs. Half
3.8 Siblingship – Full vs. Unrelated
3.9 Siblingship – Half vs. Unrelated
3.10 Example Report Siblingship - Full vs. Half
4.0
Y Chromosome Testing
5.0
Mitochondrial Testing
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DNA Collection and
Management
When Quality Matters
More and more customers are choosing to do
business with a company based on whether or not
they are certified in their particular area of
expertise.
A well trained, certified staff can offer better
customer service to your customers, which will lead
to increased referrals to your business.
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(520) 750-7555 - Tucson, AZ
ChromoCONNECTTM
ChromoCONNECT™ is your secure on-line portal to your
confidential
DNA
paternity
testing
results.
With
ChromoCONNECT™ you can:




Register your paternity cases online
Monitor the status of your DNA test as it moves through
the laboratory
Download and print your DNA test report when it’s
complete.
Find collection locations in other cities/states
How do I get started?
Go to www.chromosomal-labs.com and click on the “affiliates”
button, then click on “ChromoCONNECT”. Here you will be able
to sign up to use ChromoCONNECT™. After we receive your
information we will set up your account and then contact you with
instructions on how to use it.
What does it mean to register a case?
After logging into your account you can fill out your sample information (Chain of Custody) online. After you have filled out all of
the information, you will print the chain of custody form and submit it along with your samples.
What is the benefit of using ChromoCONNECT™?
ChromoCONNECT™ was designed to flag missing information prior to submission of the case. When a case is submitted with
missing information it can delay the release of your case by one or more days. ChromoCONNECT™ ensures all the information
has been entered properly resulting in a timely release of your case without administrative delays. Another benefit is because
all the information is typed into the system, all the information is legible. With legible information, all names on the report will be
100% accurate, whereas there can be errors in the name spellings when the information is illegible.
How do I monitor my case status online?
Once your samples are received at our laboratory they are scanned into ChromoCONNECT™. You can login to your account
where you can view a list of all your cases. By clicking on a particular case you will be able to see:




when your samples went into the lab for analysis
when your report is in the review process
when results are complete and ready to print
when hardcopies have been shipped
If there is a delay in the analysis for any reason, it will be noted, along with the reason and a new date to expect results.
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(520) 750-7555 - Tucson, AZ
ChromoCONNECTTM
What do I do if my customer(s) live in a different city or state?
You can use ChromoCONNECT™ to find another collector to perform the collection for you. We refer up to 2 locations per
city. Collectors who are certified by NADCM (www.NADCM.com) are referred first to ensure you are provided with the most
qualified collectors.
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(520) 750-7555 - Tucson, AZ
DNA Immigration Testing
Technical Bulletin 40-006.1 - DTC
United States citizens can petition for their
spouses, parents, children or siblings to immigrate
to the United States and permanent residents can
do so for their spouses and unmarried children.
In most cases there are sufficient written records to
validate relationships and satisfy the requirements
of the U.S. Citizen and Immigration Services
Department. If more evidence of a biological
relationship is needed, a voluntary DNA test may
be advised. Tests are typically requested for
immigration and passports and sometimes for
other reasons. It is important to ask the reason for
testing, as that could dictate which U.S. agency is
responsible for sample collection and receiving
reports.
Test Fees
Fees must be paid by the test participants. The
fees are not paid by the USCIS or Embassy. There
are two fees associated with Immigration DNA
testing, the DNA test fee and an immigration
handling fee.
DNA Collection of Petitioner
The person residing in the United States, the petitioner will be collected at a qualified laboratory approved
location. They will need a government issued photo identification and a copy of a letter from the USCIS/
Embassy asking for a DNA test. If this letter is not available we need any document they have received
stating their government issued case number. This paperwork will allow the USCIS/ Embassy to match the
laboratory results to the correct government case. This letter, a photo taken at the time of collection and a
photocopy of the government issued photo ID must accompany the samples to Chromosomal Labs ● Bode
Technology.
DNA Collection of Beneficiary
The person outside of the United States, the beneficiary, will be collected at the U.S. Embassy in that
country or at a site approved by the U.S. Embassy. A sample collection kit must be sent from Chromosomal
Labs ● Bode Technology to the foreign location. This shipping cost is included in the immigration handling
fee. Chromosomal Labs ● Bode Technology will supply you with the tracking number, or the appointment
Page 1 of 2
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DNA Immigration Testing
Technical Bulletin 40-006.1 - DTC
information. Some countries contact the persons in their country with an appointment time, while other
countries contact the petitioner with an appointment time. On the other hand, some countries contact the
laboratory with an appointment time. The fees to be paid for the collection are NOT included in the
immigration fee. The cost is bore by the person(s) in the foreign country. The beneficiary needs to take
government issued identification with them to the collection appointment.
Shipping Samples from a Foreign Country
After collection, the samples are sent directly to Chromosomal Labs ● Bode Technology by the U.S.
Embassy or their designee. The fees for this shipment are included in the immigration handling fee. The
laboratory will provide you with a tracking number for this shipment; however we cannot guarantee that the
shipper will use the air bill provided. Please note, a small number of countries require the person(s) in the
foreign country to bear these shipping costs. In this instance, the immigration handling fee is prorated
accordingly. The laboratory will notify you if you are dealing with one of these countries.
DNA Testing
Once all of the samples and necessary documentation have been received, a sample receipt email will be
sent to inform you that the samples have arrived from the foreign country. Test results will be available in 2-3
business days, depending on the testing scenario. Initially you will receive the test results by fax or email.
Final Report
The U.S. Government requires that the final results be received directly from the testing laboratory.
Chromosomal Labs ● Bode Technology will ship two notarized copies of the test results and the original
photographs and documents to the USCIS office or U.S. Embassy. It is important that the test participants
ensure the laboratory knows where to ship the results. Chromosomal Labs ● Bode Technology will provide a
FedEx tracking number for this shipment. The fees for this shipment will be included in the immigration
handling fee.
Follow Up
The USCIS typically requires one to two weeks to process once they have received the result package from
Chromosomal Labs ● Bode Technology.
Page 2 of 2
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MiniFiler for Degraded or
Inhibited Samples
DNA Degradation
The quality of DNA samples collected from a crime scene are often less
than ideal, having been exposed to harsh environmental conditions for
prolonged periods of time. As a consequence of exposure to water and
heat, DNA degrades into small fragments from a number of bacterial,
biochemical or oxidative processes.
Successful analysis of degraded DNA from compromised forensic
evidence improves considerably with smaller-sized amplicons.
Unfortunately, amplicon size and the ability to amplify extremely
degraded DNA were not considered during the commercial development
of the multiplex kits that incorporate the 13 CODIS loci. Consequently,
several of these loci have a large number of repeat units or wide allele
ranges that are not favorable for generating small amplicons.
PCR Inhibition
Biological fluids from crime scenes are often associated with soil, sand, wood, textile dyes, leather or other
substrates that contain materials which may co-extract with the DNA and prevent PCR amplification. Samples
containing PCR inhibitors often produce partial DNA profiles that appear similar to samples that contain
degraded DNA.
MiniFiler™ Kit
The MiniFiler Kit is the world's first commercially available miniSTR kit and it substantially increases the ability
to obtain DNA results from compromised samples that previously would have yielded limited or no genetic
data.
The MiniFiler™ Kit tests for the following nine challenging CODIS loci: D13S317, D7S820, Amelogenin,
D2S1338, D21S11, D16S539, D18S51, CSF1PO, and FGA and compliments other autosomal STR kits that
are currently being used for forensic casework. Loci that drop out using conventional STR kit technology may
be recovered using the MiniFiler™ Kit. The kit also combines a proprietary buffer with optimized thermocycling
parameters to enable the system to overcome inhibitors commonly encountered in forensic samples
The MiniFiler™ Kit is an extremely valuable tool in casework that originally produced minimal to no DNA
results, missing persons identification and disaster victim identification. Use of a dual-amplification strategy
using MiniFiler™ and Identifiler™ Kits is the best strategy for recovery of all 15 autosomal markers from
compromised samples.
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Keeping Current with Cutting Edge Technologies
DNA profiling has emerged over the past
thirty years as a technique for the
identification
of
individuals.
Early
techniques such as RFLP, Southern
Blot, HLA-DQ-Alpha and Polymarker, all
served a purpose in the evolution of
DNA identification, but the lack of
discriminating power and the frequent
inability to resolve mixtures resulted in
the desire for better methods. The
invention of the Polymerase Chain
Reaction (PCR) Technique allowed for
substantial increases in discriminating
power, the ability to recover profiles from
degraded samples, and the ability to
recover useable information from very
small samples. Given a suitable number
of discrete markers, it is now possible to
distinguish anyone on the planet with the
exception of identical twins. Although
rudimentary in the beginning, continual
research
and
development
has
constantly added new techniques and
new marker systems to relentlessly
improve the power of discrimination.
The vast majority of human identification laboratories today rely upon an adaptation of the PCR method combined
with short tandem repeat (STR) analysis. STR methods use highly polymorphic regions that contain short repeated
sequences of DNA. Since unrelated individuals will often have different numbers of repeat blocks, and the analysis
incorporates fifteen (15) independently inherited loci, discriminating power increases exponentially.
Raising the Bar: Providing the Highest Quality Results
In an effort to provide the best possible results, Chromosomal Labs ● Bode Technology embraces a concept of
continual improvement in services of value to our customer base. Our basic platform, Identifiler Plus is the most
discriminating kit available for commercial use. This collection of 15 (plus Amelogenin) markers works extremely well
in 99% of the 60,000 relationship cases our laboratory performs annually. For these cases, comparison of mother
and child results in a known set of obligate alleles that must come from the father.
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For cases where it is necessary to establish a relationship on a more extended family group, adding a more highpowered marker system such as NGM can be extremely valuable. Full and half siblingships, avuncular (aunts and
uncles), grandparentage and family reconstructions will at times result in inconclusive results. By employing extra
markers these relationships have a much higher degree of relationship resolution.
The NGM PCR Amplification Kit contains 15 autosomal short tandem repeat (STR) loci. An overlap exists of ten
markers already appearing in the Identifiler kit, but the additional 5 markers are highly polymorphic which results in a
high degree of usefulness in our cases. The new markers, which have been in use for years in Europe, are:
 D1S1656
 D2S441
 D10S1248
 D12S391(appears on the 12th chromosome with vWA)
 D22S1045
Twenty-Eight Markers Available to Resolve Difficult Relationship Puzzles
When we consider other markers we have added previously, namely Penta E, Penta D, and the set of 5 markers
from CS7, a 28 marker system is now available. Special notice is given to D12S391 and vWA, Penta E and FESFPS
as their recombination distance confirms a linkage where independence cannot be assumed for kinship analysis.
Exceptional rules are employed to provide the highest accuracy in kinship analysis. The current convention for
relationship testing allows one or at times two single step mismatches to be considered mutations which can lead to
an inconclusive result. Adding these five (5) very discriminating markers will often separate true mutations from
exclusions.
When required, our ability to deploy the NGM markers has shown the ability to increase our Combined Paternity
Index in cases where the relationship is established. In other cases where there is a low probability of relatedness,
combined likelihood ratios (CLR) have been lowered, as expected.
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Infidelity Testing
Sexual infidelity is a serious problem
plaguing relationships today and is
associated with approximately seventeen
percent of all divorces in the United States.
While estimates of the frequency of marital
infidelity vary widely, one well-known expert,
Peggy Vaughan, author of The Monogamy
Myth, estimates that 60 percent of husbands
and 40 percent of wives will have an affair at
some point during their marriage.
Investigation of Female Infidelity
The presence of stains on female undergarments can be a possible indicator of sexual infidelity with a male. During
and after sexual activity, semen can be deposited on undergarments, panties, bed sheets, clothing, upholstery or
other surfaces. Sperm cells can reside in the vagina for up to five days after intercourse and can remain identifiable
for years in dried stains. Stains on female undergarments can also be attributable to urine, urinary tract infection,
blood, feces or natural vaginal excretions associated with menstruation, ovulation or vaginal infection.
Laboratory analysis of suspicious stains is performed in two stages, screening and DNA analysis.
In the first stage, the suspicious stain is screened by a Forensic Analyst for the presence of semen using a series of
techniques.
If semen is detected in the stain, DNA analysis of the stain and a reference swab sample from the partner is
performed to determine if the semen belongs to the partner or another unknown male. Biological stains resulting
from sexual activity often contain a mixture of both male and female DNA. The female DNA usually originates from
epithelial cells from the vaginal wall, the mouth or skin. Stains are first processed with a differential extraction that
takes advantage of the unique characteristic of each cell type. A DNA profile is generated from each stain extract
and DNA from the male reference. If the male DNA in the stain does not match that of the DNA from the reference
man, the results indicate that the semen stain is from another man.
Investigation of Male Infidelity with a Female
The presence of stains on male undergarments can be a possible indicator of sexual infidelity with a female. Natural
staining of male undergarments can be caused by urine, urinary tract infection, feces or semen. Similarly, stains to
undergarments from sexual infidelity with a female can be caused by semen, vaginal fluids, blood, saliva or feces.
Laboratory analysis of suspicious stains is typically performed in two stages, screening and DNA analysis.
In the first stage, the suspicious stain is screened by a Forensic Analyst for the presence of semen, saliva or other
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Infidelity Testing
biological material using a series of techniques. After screening, DNA analysis of the stain and a reference swab
sample from the known partner is performed. Biological stains resulting from sexual activity often contain a mixture
of both male and female DNA. The female DNA usually originates from epithelial cells from the vaginal wall, the
mouth or skin.
Stains are first processed with a differential extraction that takes advantage of the unique
characteristic of each cell type. A DNA profile is generated from each stain extract and from the female reference.
If the female DNA in the stain does not match the DNA from the reference female, the results indicate the presence
of another woman.
Investigation of Male Infidelity with a Male
The presence of stains on male undergarments can be a possible indicator of sexual infidelity with a male. Natural
staining of male undergarments can be caused by urine, urinary tract infection, feces or semen. Similarly, stains to
undergarments from sexual infidelity with a male can be caused by semen, saliva, blood or feces.
Laboratory analysis of suspicious stains is typically performed in two stages, screening and DNA analysis.
In the first stage, the suspicious stain is screened by a Forensic Analyst for the presence of semen, saliva or other
biological fluids using a series of techniques. After screening, DNA analysis of the stain and a reference swab
sample from the known partner is performed. Biological stains from homosexual activity often contain a mixture of
DNA and can originate from epithelial cells from the anus, the mouth or skin. Stains are first processed with a
differential extraction that takes advantage of the unique characteristic of each cell type. A DNA profile is generated
from each stain extract and from the male reference. If the male DNA in the stain does not match the DNA from the
reference male, the results indicate the presence of another man.
Limitations of Infidelity Testing

Condom use often precludes the presence of semen for testing; however it may or may not eliminate foreign
DNA from saliva or other sources.
General Guidelines for Sample Collection


Garments and other evidence samples should be placed into paper bags or envelopes and stored at room
temperature. Samples should not be placed in plastic bags or refrigerated.
The reference sample from the known partner should be collected from the inside of the cheek using sterile
swabs provided by the laboratory.
Summary of Semen Screening Methods
Suspicious stains can be rapidly and reliably screened for the presence of semen utilizing a combination of three
techniques: ultraviolet illumination, prostrate specific antigen (PSA) and microscopy.
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Infidelity Testing
Ultraviolet Examination
Ultraviolet (UV) light is a popular tool in many forensic investigations for the presumptive identification of body
fluids on a variety of substrates. Materials such as semen, saliva, perspiration and vaginal secretions are
naturally fluorescent under UV light which offers a discriminating method for locating these stains which might
otherwise be invisible to the naked eye. Once the precise location of the stain is determined, confirmatory
testing can be conducted utilizing the PSA and microscopy techniques.
Prostate Specific Antigen
Prostate-specific antigen, PSA, is a glycoprotein produced in the prostate and secreted into seminal fluid.
PSA is one of the major proteins in seminal fluid with concentrations of 0.2 to 3.0 mg/ml. In sharp contrast,
PSA is found in very low concentrations in vaginal fluid, ranging from 0 to 1.25 ng/ml. This makes PSA a
useful forensic marker for the detection of small amounts of seminal fluid. It can be found in the absence of
spermatozoa in the case of vasectomized men. PSA demonstrates good stability and on average is
detectable in vaginal smears 27 hours after intercourse. PSA has been recovered from semen stains as old
as 30 years.
Microscopy
Sperm heads can be accurately identified based on their morphological characteristics via microscopy.
Summary Typical Result Scenarios for Semen Screen
Semen Screen Test Result
PSA
Sperm
Negative
Negative
Negative
Positive
Positive
Positive
Positive
Positive
Positive
Negative
Conclusion
No evidence of semen
Presence of semen confirmed
Presumptive evidence of semen
44
Comments
DNA testing is necessary. PSA is a strong
indicator of the presence of semen, however
it can be found in low concentrations in
vaginal sections. This type of result is also
typical in men that have been vasectomized.
(520) 750-7555 - Tucson, AZ

DNA Testing Handbook - Advantage Medical Products

Transcription

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