Simoa Total PSA ™ USER-101-01

Simoa™ Total PSA
USER-101-01
11 Feb 2014
Quanterix Corporation
113 Hartwell Avenue
Lexington, MA 02421
Note: Read highlighted exceptions
that apply to Simoa software version 1.1 users
Customer Support
1-877-786-8749
[email protected]
For Research Use Only. Not for use in diagnostic procedures.
Read this package insert prior to use. Package insert instructions must be
carefully followed. Reliability of assay results cannot be assured if there are
deviations from the instructions in this package insert.
NAME
Simoa™ Total PSA
INTENDED USE
The Simoa Total PSA assay is a digital immunoassay for the quantitative
determination of low-abundance total PSA in human serum.
SUMMARY AND EXPLANATION OF TEST
Prostate specific antigen (PSA) is a serine protease with chymotrypsin-like
activity. It is a member of the kallikrein-related peptidase gene family. PSA is a
single-chain glycoprotein of 237 amino acids with a molecular weight of
approximately 30,000 daltons. PSA contains 7%–8% carbohydrate as a single
N-linked oligosaccharide side chain.1 The major site of PSA production is the
glandular epithelium of the prostate, but it has also been found in breast
cancers, salivary gland neoplasms, periurethral and anal glands, cells of the
male urethra, breast milk, blood, and urine. 1,2 PSA produced in the prostate is
secreted into the seminal fluid, where its major function is the proteolytic
cleavage of gel-forming proteins in the seminal fluid, resulting in the
1
liquification of the seminal gel and increased sperm mobility. Small quantities
of PSA are found in the blood of men as a result of leakage of PSA from the
prostate gland. Increasing levels of serum PSA are associated with prostatic
pathology, including prostatitis, benign prostatic hyperplasia (BPH), and cancer
of the prostate.1, 3–7
PSA occurs in two major immunodetectable forms in blood. The major form is
PSA complexed with the serine protease inhibitor, -1-antichymotrypsin
(PSA-ACT). Uncomplexed, or free, PSA is the other detectable form of PSA in
serum. The majority of free PSA in serum appears to be an inactive form that
cannot complex with protease inhibitors and may be either a PSA zymogen or
an enzymatically inactive cleaved form of PSA. Equimolar-response PSA assays
1
have an equivalent response to both free PSA and PSA-ACT. The Simoa Total
PSA assay uses reagents that recognize both free PSA and PSA-ACT equally.
Measurement of PSA following radical prostatectomy (RP) has become
standard practice for prostate cancer recurrence monitoring. Biochemical
8–11
cancer relapse occurs in up to 40% of patients after RP,
and a third of these
patients develop metastatic disease with a 20% probability of dying within 10
12,13
years.
PSA is typically undetectable by most assay methods following
surgery, and it is generally agreed that undetectable post-surgical PSA over
time is indicative of a good prognosis.14–16 However, low-abundance PSA could
be rising while remaining below the detection limits of conventional
immunoassays, and consideration of potential salvage therapies awaits the
emergence of PSA above a reliably quantifiable threshold, generally 0.1 or 0.2
ng/mL. Clinical data indicate that early adjuvant and salvage radiation therapies
17–20
following surgery significantly improve patient outcomes.
Recent clinical
data with extreme sensitivity, non-conventional immunoassay (immunoPCR
and digital immunoassay) indicate potential utility from low-abundance PSA
measurement following RP for risk stratification and early cancer recurrence
monitoring.21–24
PRINCIPLES OF THE PROCEDURE
The Simoa Total PSA assay is a 3-step digital immunoassay to measure the
quantity of total PSA in human serum using the Simoa HD-1 Analyzer and Single
Molecule Array (Simoa) technology. 25
In the first step, anti-PSA coated paramagnetic capture beads are combined
with sample. PSA molecules present in the sample are captured by the anti-PSA
coated capture beads. After washing, biotinylated detector antibodies are
mixed with the capture beads. The detector antibodies bind to the captured
PSA. Following a second wash, a conjugate of streptavidin-ß-galactosidase
(SBG) is mixed with the capture beads. SBG binds to the biotinylated detector
antibodies, resulting in enzyme labeling of captured PSA. Following a third
wash, the capture beads are resuspended in a resorufin ß-D-galactopyranoside
(RGP) substrate solution and transferred to the Simoa Disc. Individual capture
beads are then sealed within microwells in the array. If PSA has been captured
and labeled, the ß-galactosidase hydrolyzes the RGP substrate into a
fluorescent product that provides the signal for measurement. A single labeled
PSA molecule results in sufficient fluorescent signal in 30 seconds to be
detected and counted by the Simoa optical system. At low PSA concentration,
the percentage of bead-containing wells in the array that have a positive signal
Simoa
TM
Total PSA
is proportional to the amount of PSA present in the sample. At higher PSA
concentration, when most of the bead-containing wells have one or more
labeled PSA molecules, the total fluorescence signal is proportional to the
amount of PSA present in the sample. The concentration of PSA in unknown
samples is interpolated from a standard curve.
For additional information on system and assay technology, refer to the Simoa
HD-1 Analyzer User Guide.
REAGENTS
Reagent Kit
Simoa Total PSA kit (101)
Bead Reagent
1 bottle
(13.1 mL)
Detector Reagent
1 bottle
(13.1 mL)
SBG Reagent
1 bottle
(13.1 mL)
RGP Reagent
PSA Calibrator
Concentrate
2 bottles
(3.4 mL)
2 bottles
(0.2 mL)
PSA Calibrator
Diluent
2 bottles
(9.5 mL)
Sample Diluent
1 bottle
(10.7 mL)
Anti-PSA (mouse monoclonal) coated
capture beads in Tris buffer with protein
stabilizers (bovine). Preservative: ProClin
300.
Biotinylated anti-PSA (mouse
monoclonal) in phosphate buffer with
protein stabilizers (bovine). Preservative:
ProClin 300.
Conjugate of streptavidin-ß-galactosidase
(SBG) in phosphate buffer with protein
stabilizers (bovine). Preservative: ProClin
300.
Resorufin ß-D-galactopyranoside (RGP) in
phosphate buffer with a surfactant.
PSA antigen in phosphate buffer with
protein stabilizers (bovine). Preservative:
ProClin 300.
Phosphate buffer with bovine serum
components and a surfactant.
Preservative: ProClin 300.
Phosphate buffer with bovine serum
components, a heterophilic blocker, and a
surfactant. Preservative: ProClin 300.
Other Reagents
System Wash Buffer 1 (ordered separately)
Phosphate buffer with surfactant. Preservative: ProClin 300.
System Wash Buffer 2 (ordered separately)
Phosphate buffer. Preservative: ProClin 300.
Sealing Oil (ordered separately)
Synthetic fluorinated polymer.
WARNINGS AND PRECAUTIONS
For research use only. Not for use in diagnostic procedures.
Safety Precautions
CAUTION: This product requires the handling of human specimens. It is
recommended that all human-sourced materials be considered potentially
infectious and be handled in accordance with the OSHA Standard on
26
27
Bloodborne Pathogens. Biosafety Level 2 or other appropriate biosafety
28
practices should be used for materials that contain or are suspected of
containing infectious agents.
The Simoa Total PSA reagents contain methylisothiazolones, which are
components of ProClin and are classified per applicable European
Community (EC) Directives as: Irritant (Xi). The following are the appropriate
Risk (R) and Safety (S) phrases.
R43 May cause sensitization by skin contact.
S24 Avoid contact with skin.
S35 This material and its container must be disposed of in a safe way.
S37 Wear suitable gloves.
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S46 If swallowed, seek medical advice immediately and show this container
or label.
For a detailed discussion of safety precautions during instrument operation,
refer to the Simoa HD-1 Analyzer User Guide.
Package insert instructions must be carefully followed. Reliability of assay
results cannot be assured if there are deviations from the instructions in this
package insert.
Handling Precautions
Specimens may be stored for up to 24 hours at 2–8°C prior to being tested. If
testing will be delayed for more than 24 hours, specimens should be frozen
at –20°C or colder.
Failure to follow these instructions may result in inaccurate or inconsistent
specimen results.
The Simoa HD-1 Analyzer does not provide the capability to verify specimen
type. It is the responsibility of the operator to verify that the correct
specimen type is used in the Simoa Total PSA assay.
Do not use reagent kits beyond the expiration date.
Use caution when handling patient specimens to prevent crosscontamination. Use of disposable pipettes or pipette tips is recommended.
Do not pool reagents within a kit or between reagent kits.
Do not use grossly hemolyzed specimens.
Do not attempt to reuse cuvettes or Simoa Discs, as this will cause significant data quality deterioration.
For optimal results, inspect all samples for bubbles. Remove bubbles with an
applicator stick prior to analysis. Use a new applicator stick for each sample
to prevent cross-contamination.
Prior to loading the Simoa Total PSA reagents on the Simoa HD-1 Analyzer
for the first time, the Bead Reagent bottle must be mixed to resuspend
capture beads that have settled during shipment. For capture bead–mixing
instructions, refer to the Procedure section of this package insert.
For a detailed discussion of handling precautions during instrument
operation, refer to the Simoa HD-1 Analyzer User Guide.
Storage Instructions
Multiple freeze-thaw cycles of specimens should be avoided.
Specimens with obvious microbial contamination should not be used.
Specimen stability in different storage conditions has not been validated for
this assay.
PROCEDURE
Materials Provided
Simoa Total PSA Reagent Kit
Simoa Total PSA reagents must be stored at 2–8°C in an upright
position and may be used immediately after removal from 2–8°C storage.
PSA Calibrator Concentrate bottles must be stored at –20°C.
When stored and handled as directed, reagents and PSA Calibrator
Concentrate are stable until the expiration date.
The Simoa Total PSA reagents should be removed from the instrument upon
completion of the assay run and stored at 2–8°C upright with caps on.
Indications of Reagent Deterioration
When a user-prepared QC control value is out of its expected range, it may
indicate deterioration of the reagents or errors in technique. Associated test
results may be invalid and may require retesting. Assay recalibration may be
necessary. See also the Quality Control Procedures section of this package
insert.
SPECIMEN COLLECTION AND PREPARATION FOR
ANALYSIS
Insufficient sample processing may cause inaccurate results.
For optimal results, serum specimens should be free of fibrin, red blood
cells, or other particulate matter. Centrifuge specimens containing fibrin, red
blood cells, or particulate matter prior to use to ensure consistency in the
results.
Note that interfering levels of fibrin may be present in samples that do not
have obvious or visible particulate matter.
Specimens thawed after frozen storage must always be mixed THOROUGHLY
by low-speed vortexing or inverting 10 times. Visually inspect the specimens.
If layering or stratification is observed, continue mixing until specimens are
visibly homogeneous. Centrifuge the specimens prior to assay.
Centrifugation conditions should be sufficient to efficiently remove
particulate matter and to clarify the sample.
Centrifuged specimens with a lipid layer on the top should be transferred to
a secondary tube. Care must be taken to transfer only the clarified specimen
without the lipemic material.
For freshly drawn serum specimens, ensure that complete clot formation
has taken place prior to centrifugation. If specimens are centrifuged before a
complete clot forms, the presence of fibrin or particulate matter may cause
erroneous results.
Serum should be immediately removed from the red cells (after
centrifugation) and put in a separate tube that can then be aliquoted and
frozen for future use.
Simoa
TM
Total PSA
Materials Required But Not Provided
Simoa HD-1 Analyzer
Simoa HD-1 System Wash Buffer 1
Simoa HD-1 System Wash Buffer 2
Simoa HD-1 Sealing Oil
Simoa cuvettes
Simoa disposable pipettor tips
Simoa Discs
For information on materials required for maintenance procedures, refer to
the Simoa HD-1 User Guide.
Instrument Procedure
The Simoa Total PSA assay definition must be installed on the Simoa HD-1
Analyzer prior to performing the assay. For information on obtaining and
installing the assay definition, refer to the Simoa HD-1 Analyzer User Guide.
Assay Procedure
Prepare calibrators per the Preparing Calibrators section of this package
insert. If desired, prepared QC controls per the Preparing QC Controls
section of this package insert.
Reagents must be allowed to come to room temperature before loading
onto the HD-1 Analyzer.
Before loading the Simoa Total PSA reagents on the Simoa HD-1 Analyzer,
the Bead Reagent bottle must be mixed to resuspend capture beads that
have settled during shipping and storage. To resuspend the beads, vortex for
a minimum of 30 seconds. (Note: The bead reagent is formulated with an
antifoam agent, but vortexing can still create foaming. If the foam does not
dissipate within a few minutes, remove excess foam with a pipette prior to
loading capture beads onto the Simoa HD-1 Analyzer.)
Before loading a new bottle of RGP Reagent on the Simoa HD-1 Analyzer for
the first time, mix the reagent by inversion 10 times to ensure homogeneity
(the reagent is stored frozen prior to shipment from Quanterix™).
Set up the assay run on the instrument (see the Simoa HD-1 Analyzer User
Guide).
Load the Simoa Total PSA reagents (Bead Reagent, Detector Reagent, SBG
Reagent, Sample Diluent) into the reagent bay.
Load samples, calibrators, controls, and RGP into the sample bay. (Note:
Only one bottle of RGP is needed per 96-sample run.)
Follow the guidelines in the Specimen Dilution Procedures section of this
package insert to set up sample dilutions.
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The minimum sample volume needed depends on the number of
replicates desired, the pipetting protocol chosen (standard or neat), and
whether samples will be introduced to the instrument in tubes or
Quanterix-supplied 96-well plates. If a sample tube is used, the type and
volume of the tube may determine the dead volume. As a general
guideline, the required sample volume may be calculated as illustrated in
the following examples:
Calculates PSA results by interpolation from a calibration curve.
For information on setting up assay runs, data analysis, and general operating
procedures, refer to the Simoa HD-1 Analyzer User Guide.
Samples introduced in 5-mL Nalgene Cryogenic sample tubes:
Calculation: 50 µL (dead volume of tube) + (number of replicates ×
protocol aspiration volume)
Example: (PSA neat protocol) 100 µL of sample are aspirated. Three
replicates are desired. Required sample volume = 50 µL + (3 × 100 µL) =
350 µL.
Samples introduced in a Quanterix-supplied 96-well plate:
Calculation: 50 µL (dead volume of well) + (number of replicates ×
protocol aspiration volume)
Example: (PSA standard protocol) 25 µL of sample are aspirated. Two
replicates are desired. Required sample volume = 50 µL + (2 × 25 µL) =
100 µL. Note: The maximum recommended sample volume for the
Quanterix-supplied 96-well plates is 175 µL. When assaying calibrators
from 96-well plates, each replicate of calibrator must be placed in a
separate well due to volume limitations (100-µL protocol aspiration
volume + 50-µL well dead volume). Note: The PSA standard protocol is
unavailable with Simoa software version 1.1. The available protocol
corresponds to the neat protocol described in this section.
For optimal performance, it is critical to follow the routine maintenance
procedures defined in the Simoa HD-1 Analyzer User Guide.
Note: If samples are left on board the instrument or pipetted samples are
left on a lab bench for more than an hour, evaporation effects will
influence the results. The effect depends on the volume of sample. As a
general guideline, at room temperature and normal humidity, a 60-µL
sample will lose approximately 5% of its weight per hour. A 120-µL
sample may lose 5% of its weight in approximately 3 hours.
Replenish consumables and system resources as needed prior to initiating
the run, as described in the Simoa HD-1 Analyzer User Guide.
Initiate the run.
During the run the Simoa HD-1 Analyzer performs the following functions:
Primes washers and sealing oil tubing and washes the fixed tip pipettor.
Adds cuvettes to the incubation ring.
Adds capture beads to a cuvette.
Pellets capture beads with a magnet and removes bead diluent.
Adds sample to the cuvette and shakes the cuvette to mix beads and
sample.
Incubates the capture bead–sample mixture.
Moves the cuvette to a wash station and washes capture beads with System
Wash Buffer 1.
Removes System Wash Buffer 1 from the cuvette and adds Detector
Reagent, followed by shaking and incubation.
Moves the cuvette to a wash station and washes capture beads with System
Wash Buffer 1.
Removes System Wash Buffer 1 from the cuvette and adds SBG Reagent,
followed by shaking and incubation.
Note: Simoa software version 1.1 users are required to use off-line analysis
software for generated results. Refer to the training instructions provided
during instrument installation.
Specimen Dilution Procedures
Specimens with a PSA value exceeding 400 pg/mL (PSA standard protocol,
1:4 specimen pre-dilution) are flagged with an error code. Note: The PSA
standard protocol is unavailable with Simoa software version 1.1. Version
1.1 is capable of running only the neat protocol, which does not perform a
1:4 specimen pre-dilution. Therefore, results in 1.1 will be flagged with an
error code if specimen PSA value exceeds 100 pg/mL.
To request sample dilutions by the instrument, the operator must enter the
desired dilution factor in the instrument software set-up screen. All samples
selected for dilution in that run will be diluted. (Note: if different singleplex
assays are selected for a given sample, different dilutions can be selected for
each assay). The system will use this dilution factor to automatically
calculate the concentration of the sample before dilution and report the
result. The dilution should be performed so that the diluted result reads
within the calibration curve range. Note: This section does not apply to users
of Simoa software version 1.1. The 1.1 software does not perform sample
dilutions. In this case, users must manually perform 1:4 pre-dilutions of
samples with the provided Sample Diluent prior to running on the Simoa
HD-1 Analyzer. Pre-dilution with Sample Diluent is required to introduce
heterophilic blocking activity to the assay.
For detailed information on ordering dilutions, refer to the Simoa HD-1
Analyzer User Guide.
Preparing Calibrators
For maximum accuracy in preparing calibrators, it is recommended that PSA
Calibrator Concentrate and PSA Calibrator Diluent be brought to room
temperature prior to pipetting. Thaw a vial of frozen PSA Calibrator
Concentrate (approximately 1 hour at room temperature). Do not heat the
vial to accelerate thawing. When the solution is fully thawed, THOROUGHLY
mix by multiple gentle inversions or vortexing. Frozen protein solutions can
partition during freezing, so complete mixing of thawed material is critical
for accurate calibrators.
Prepare 1000 pg/mL stock solution (depicted in the diagram below) from the
Calibrator Concentrate. Refer to the value assignment for the lot of PSA
Calibrator Concentrate being used to prepare the 1000 pg/mL stock solution.
Prepare 8 calibrators by 1:3 serial dilution. Pipette 900 µL of PSA Calibrator
Diluent into the 100 pg/mL tube (see diagram below). Pipette 667 µL of
diluent into each remaining tube. Pipette 100 µL of the PSA Calibrator
Concentrate (1000 pg/mL) into the 100 pg/mL tube and vortex. Transfer 333
µL of the 100 pg/mL tube into the 33.3 pg/mL tube. Repeat the series,
mixing each tube thoroughly before the next transfer. Use PSA Calibrator
Diluent as the zero calibrator.
Moves the cuvette to a wash station and washes capture beads with System
Wash Buffer 1. The final wash uses System Wash Buffer 2 to remove
surfactant.
Removes System Wash Buffer 2 from the cuvette and adds RGP Reagent.
Transfers the capture bead mixture to the inlet port of a
Simoa Disc.
Applies negative pressure to the flow cell from a ventilation port in the disc,
drawing the bead mixture across the readout array. Beads settle into wells
of the array.
Indexes the Simoa Disc to the sealing oil station and pipettes oil into the
inlet port of the disc. The oil seals the wells of the array.
Indexes the Simoa Disc to the imaging station and interrogates the array for
the presence of fluorescence signal growth in each well of the array.
Analyzes the array images to determine the average enzymes/bead (AEB).
Simoa
TM
Total PSA
Note: The above procedure is recommended for preparation of calibrators.
If an alternative procedure is followed, volume transfers should be no less
than 10 µL to achieve the best accuracy.
Page 4
Preparing QC Controls
To prepare QC controls, use the PSA Calibrator Concentrate and the PSA
Calibrator Diluent. It is recommended that controls be minimally placed at
approximately 3.0 and 60 pg/mL to monitor the digital and analog ranges of
the assay.
Note: Due to the sensitivity of the Simoa Total PSA assay, PSA is detectable
in female serum.
Calibration and Dynamic Range
Calibration should be performed with multiple replicates of each calibrator
level (two or three replicates).
Calibration curve validity should be assessed by assaying QC controls. Ensure
that control values are within the concentration ranges expected based on
QC value assignment and control range determination testing protocols in
use in your laboratory.
The Simoa HD-1 Analyzer PSA standard pipetting protocol automatically
performs a 4-fold pre-dilution of unknown PSA samples, but calibrators are
not pre-diluted. This 1:4 pre-dilution leverages the high sensitivity of Simoa
digital assays to help minimize interferences and matrix effects, while
extending the dynamic range of the assay for unknowns to 400 pg/mL. Note:
The PSA standard protocol is unavailable in Simoa software version 1.1. Only
the neat protocol is available in 1.1.
If it is desired to assay unknown PSA samples without the 1:4 pre-dilution,
the neat PSA protocol can be specified. (Note: Assaying samples without
pre-dilution with Sample Diluent may raise the risk of non-specific
interferences due to the absence of blocking agents formulated into the
Sample Diluent.)
Once a Simoa Total PSA calibration is stored, all subsequent samples may be
tested without further calibration unless:
A reagent kit with a new lot number is used.
QC controls are out of their expected range.
Note: Curve storage has not been qualified for Simoa software version 1.1
users.
For detailed information on how to perform an assay calibration, refer to the
Simoa HD-1 Analyzer User Guide.
Calibration Curve Fitting
The Simoa Total PSA assay uses a 4 Parameter Logistic Curve fit data reduction
2
method (4PLC, 1/y weighted) to generate a calibration curve.
Flags
Some results may contain information in the Flags field. For a description of the
flags that may appear in this field, refer to the Simoa HD-1 Analyzer User Guide.
QUALITY CONTROL PROCEDURES
It is recommended to run at least one QC control with every batch run to assess
calibration curve storage integrity and run validity. If the results from one or
more of the controls are outside the expected range for that control (based on
the QC value-assignment and range-setting procedures in your laboratory), the
stored calibration curve may no longer be valid, and the assay may need to be
re-calibrated. The length of time calibration curves may be stored must be
validated for each laboratory. Alternatively, calibration can be performed with
every batch run. It is recommended that QC controls be run with every
calibration to assess calibration accuracy and run validity. Note: Curve storage
has not been qualified for Simoa software version 1.1 users.
LIMITATIONS OF THE PROCEDURE
For research use only. Not for use in diagnostic procedures.
Specimens from people who have received preparations of mouse
monoclonal antibodies may contain human anti-mouse antibodies (HAMA).
Such specimens may show either falsely elevated or falsely depressed values
when tested with assay kits that employ mouse monoclonal antibodies.
Simoa Total PSA reagents contain a component that reduces the effect of
HAMA-reactive specimens.
Simoa
TM
Total PSA
Heterophilic antibodies in human serum can react with reagent
immunoglobulins, interfering with immunoassays. People routinely exposed
to animals or animal serum products can be prone to this interference, and
anomalous values may be observed.
The concentration of PSA in a given specimen, determined with assays from
different manufacturers, can vary due to differences in assay methods,
calibration, and reagent specificity.
BIBLIOGRAPHY
1 McCormack RT, Rittenhouse HG, Finlay JA, et al. Molecular forms of
prostate-specific antigen and the human kallikrein gene family: A new era.
Urology 1995; 45:729–44.
2 Graves HCB. Nonprostatic sources of prostate-specific antigen: A steroid
hormone-dependent phenomenon? Clin Chem 1995; 41:7–9.
3 Kuriyama M, Wang MC, Papsidero LD, et al. Quantitation of
prostate-specific antigen in serum by a sensitive enzyme immunoassay.
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4 Oesterling JE. Prostate specific antigen: A critical assessment of the most
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5 Kantoff PW, Talcott JA. The prostate specific antigen. Its use as a tumor
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6 Partin AW, Oesterling JE. The clinical usefulness of prostate specific antigen:
Update 1994. J Urol 1994; 152:1358–68.
7 Bunting S. A guide to the interpretation of serum prostate specific antigen
levels. Clin Biochem 1995; 28:221–41.
8 Gunnar A. Second-line therapy after radical prostatectomy failure: For
whom? When? How? Eur Urol 2007; 51:1155–58.
9 Bock JL, Klee GG. How sensitive is a prostate-specific antigen measurement? How sensitive does it need to be? Arch Pathol Lab Med 2004;
128:341–43.
10 Freedland SJ, Moul JW. Prostate specific antigen recurrence after definitive
therapy. J Urol 2007; 177:1985–91.
11 Han M, Partin AW, Pound CR, et al. Long-term biochemical disease-free and
cancer-specific survival
following anatomic
radical
retropubic
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12 Catalona WJ, Smith DS. 5-year tumor recurrence rates after anatomical
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13 Bianco FJ Jr, Scardino PT, Eastham JA. Radical prostatectomy: Long-term
cancer control and recovery of sexual and urinary function. Urology 2005;
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14 Oesterling JE. Prostate-specific antigen: A critical assessment of the most
useful tumor marker for adenocarcinoma of the prostate. J Urol 1991; 145:
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15 Diamandis EP. Prostate specific antigen—its usefulness in clinical medicine.
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16 Sia M, Pickles T, Morton G, et al. Salvage radiotherapy following biochemical
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Can Urol Assoc J 2008; 2:500–507.
17 Pound CR, Partin AW, Eisenberger MA, et al. Natural history of progression
after PSA elevation following radical prostatectomy. JAMA 1999;
281:1591–97.
18 Thompson IM, Tangen CM, Paradelo J, et al. Adjuvant radiotherapy for
pathological T3N0M0 prostate cancer significantly reduces risk of
metastases and improves survival: long-term followup of a randomized
clinical trial. J Urol 2009; 181:956–62.
19 Trock BJ, Han M, Freedland SJ, et al. Prostate cancer-specific survival
following salvage radiotherapy vs. observation in men with biochemical
recurrence after radical prostatectomy. JAMA 2008; 299:2760–69.
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20 Stephenson AJ, Shariat SF, Zelefsky MJ, et al. Salvage radiotherapy for
recurrent prostate cancer after radical prostatectomy. JAMA 2004;
291:1325–32.
21 Stephenson AJ, Scardino PT, Kattan MW, et al. Predicting the outcome of
salvage radiation therapy for recurrent prostate cancer after radical
prostatectomy. J Clin Oncol 2007; 25:2035–41.
22 Moul JW, Lilja H, Semmes OJ, Lance RS, Vessella RL, Fleisher M et al. NADiA
ProsVue prostate-specific antigen slope is an independent prognostic
marker for identifying men at reduced risk of clinical recurrence of prostate
cancer after radical prostatectomy. Urology 2012; 80(6):1319–25.
23 Wilson DH, Hanlon DW, Provuncher GK, Chang L, Song L, Patel PP et al.
Fifth-generation digital immunoassay for prostate-specific antigen by single
molecule array technology. Clin Chem 2011; 57:1712–21.
24 Lepor H, Cheli CD, Thiel RP, Taneja SS, Laze J, ChanDW, et al. Clinical
evaluation of a novel method for measurement of prostate-specific antigen,
AccuPSA™, as a predictor of 5-year biochemical recurrence-free survival
after radical prostatectomy: Results of a pilot study. BJU Int 2012;
109(12):1770–75.
25 Rissin DM, Kan CW, Campbell TG, et al. Single-molecule enzyme-linked
immunosorbent assay detects serum proteins at subfemtomolar
concentrations. Nat Biotech 2010; 28:595–99.
26 US Department of Health and Human Services. Biosafety in micro- biological
and biomedical laboratories, 4th ed. Washington, DC: US Government
Printing Office, May 1999.
27 World Health Organization. Laboratory biosafety manual. Geneva: World
Health Organization, 2004.
28 Clinical and Laboratory Standards Institute. Protection of laboratory workers
from occupationally acquired infections: Approved guideline, 3rd ed. CLSI
Document M29-A3. Wayne, PA: Clinical and Laboratory Standards Institute,
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Simoa and Quanterix are trademarks of Quanterix Corporation. © 2014
Quanterix Corporation. All rights reserved.
ProClin is a trademark of Rohm & Haas Company.
Nalgene is a registered trademark of Thermo Fisher Scientific or its subsidiaries.
Quanterix Corporation
113 Hartwell Avenue
Lexington, MA 02421
www.quanterix.com
Email: [email protected]
1-877-786-8749
February 2014
Part Number: 100742
Simoa
TM
Total PSA
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