Resultados Bacteriológicos (200 casos)

Resultados Bacteriológicos (200 casos)
• Positivos
• Negativos
38%
62%
Tipificación
B. Suis
B. Melitensis
B. Abortus
53%
33%
14%
SEROLOGÍA
MÉTODOS CLÁSICOS
NUEVOS MÉTODOS
HUDDLESON
ROSA DE BENGALA
AGLUT. TUBO (STA)
2ME
FIJACIÓN COMPLEM
COOMBS
BPA
ELISA INDIRECTO
C-ELISA
DIPSTICK
BRUCELLACAPT
FPA
SEROLOGIA CONVENCIONAL
•
•
•
•
•
Aglutinación placa o tubo > / = 1:160
Rosa de Bengala positiva
2 Mercaptoetanol positiva
Fijación de Complemento >/ = 1:20
Prueba de Coomb´s > >/ = 1:320
Performing the RB TEST with serum dilutions
The RB test was performed with plain serum and also with twotwo-fold
serial dilutions (from 1:1 to 1:512). The antigenic preparation was obtained
from the CVL (Weybridege, Surrey, England)
a b
c
d e f ..
white glossy
tile
1. Place 5 µl of saline in spots b, c, d, e, f…
2. Place 5 µl of patient serum in spot a anb b
3. Mix and tranfer 5 µl to b, then to c, and so on
4. Add 5 µl of RB antigen to a, b, c, etc.
Universidad
de Navarra
Universidad
de Navarra
Read the RB test with two-fold serial
dilutions in the usual way
1/2
1/4
1/8
1/16
1/32
RB titers in 126 confirmed brucellosis patients
Universidad
de Navarra
30
n = 126
Nº of sera
25
20
15
10
5
0
<4
8
16
32
64
1 2 8 2 5 6 (o r h ig h e r)
T ite rs
Titer: 88%>1:8
Usefulness of Huddleson and Rose Bengal tests for detecting antiBrucella antibodies in occupationally exposed workers from different
settings.
Jorge C. Wallach, R. A. Díaz, I. Moriyón, C. Taboadela, M.V. Delpino, P.C.
Baldi.
F.J. Muñiz Hospital ; University of San Martín; ASOCIART,
and University of Buenos Aires, Argentina; Department of
Microbiology, University of Navarra, Pamplona Spain.
Background: Although the Rose Bengal test (RB) is the screening test
recommended by WHO to detect anti-Brucella antibodies, in Argentina
local regulations recommend the use of the Huddleson test for this
purpose.
Objetive: to compare the performance of these tests for detecting
brucellosis among occupationally exposed workers in different settings
RB – Huddleson concordant cases
RB
Huddleson
SAT
Coombs
Protein
RID
8
160
160
320
negative
4
80
80
640
negative
4
128
40
640
negative
4
40
80
320
negative
128
positive
2560
>40960
positive*
4
100
80
1280
negative
1
N.D
20
1280
negative
4
25
< 20
320
negative
1
positive
80
320
negative
2
50
40
320
negative
2
25
40
40
negative
RB – Huddleson discrepant cases
RB
Huddleson
SAT
Coombs
Protein
RID
4
negative
20
640
negative
4
negative
20
1280
negative
2
negative
20
1280
negative
2
negative
20
20
negative
2
negative
20
160
negative
2
negative
20
20
negative
Conclusions
These results suggest that RB is more sensitive than Huddleson
for detecting anti-Brucella antibodies in occupationally exposed
subjects.
In discrepant cases (positive RB, negative Huddleson)
confirmatory tests (SAT and Coombs) agreed with RB.
• Desde la introducción del primer ensayo de aglutinación por Almroth Wright en 1897, una gran
variedad de métodos serológicos se han aplicado para detectar anticuerpos anti brucella.
•Si bien la seroaglutinación en tubo (SAT) sigue siendo el método standard en muchas partes del
mundo, los ensayos de ELISA han reemplazado al SAT en muchos laboratorios.
2
32
4
64
8
128
16
256
Performance of Aglutination test vs. ELISAS anti LPS and CP
Aglutination Test (%)
PAT
Acute Disease
Cronic Disease
Wright
2-ME
ELISA Test (%)
IgG a-LPS
CP
96
91
86
100
87
57
41
62
95
86
Diagnóstico Serológico
• Un título aislado de anticuerpos anti Brucella no siempre es
"diagnóstico“
• Las aglutinaciones falso negativas pueden deberse a
infección temprana o a un fenómeno prozona.
• Las reacciones falso positivas se ven infeccines cross
reactantes,Yersinia, Vibrio cholera, o por tularemia
• Los anticuerpos bloqueadores se identifican con la prueba
de Coombs.
• Cuando la aglutinación ofrece resultados poco
concluyentes deben realizarse test de Enzimo inmuno
análisis