scholarly journals Evaluation of Enzyme-Linked Immunosorbent Assay for Diagnosis of Post-Kala-Azar Dermal Leishmaniasis with Crude or Recombinant k39 Antigen

2002 ◽  
Vol 9 (2) ◽  
pp. 370-373 ◽  
Author(s):  
P. Salotra ◽  
G. Sreenivas ◽  
A. A. Nasim ◽  
B. V. Subba Raju ◽  
V. Ramesh

ABSTRACT The diagnosis of post-kala-azar dermal leishmaniasis (PKDL), a dermatosis that provides the only known reservoir for the parasite Leishmania donovani in India, remains a problem. Timely recognition and treatment of PKDL would contribute significantly to the control of kala-azar. We evaluated here the potential of the enzyme-linked immunosorbent assay (ELISA) as a diagnostic tool for PKDL. Antigen prepared from promastigotes and axenic amastigotes with parasite isolates that were derived from skin lesions of a PKDL patient gave sensitivities of 86.36 and 92%, respectively, in the 88 PKDL cases examined. The specificity of the ELISA test was examined by testing groups of patients with other skin disorders (leprosy and vitiligo) or coendemic infections (malaria and tuberculosis), as well as healthy controls from areas where this disease is endemic or is not endemic. A false-positive reaction was obtained in 14 of 144 (9.8%) of the controls with the promastigote antigen and in 14 of 145 (9.7%) of the controls with the amastigote antigen. Evaluation of the serodiagnostic potential of recombinant k39 by ELISA revealed a higher sensitivity (94.5%) and specificity (93.7%) compared to the other two antigens used. The data demonstrate that ELISA with crude or recombinant antigen k39 provides a relatively simple and less-invasive test for the reliable diagnosis of PKDL.

2019 ◽  
Vol 20 (1) ◽  
pp. 30
Author(s):  
Rinaldi Ghurafa ◽  
Denny Widaya Lukman ◽  
Hadri Latif

Brucellosis has become a zoonotic disease that received attention in efforts to prevent and eradicate strategic infectious animal diseases in Indonesia. Brucellosis can be detected early by the rose bengal test (RBT), followed by complement fixation test (CFT) and by enzyme linked immunosorbent assay (ELISA). The aims of this research was to study the indirect enzyme linked immunosorbent assay test (I-ELISA) as an alternative test for detecting brucellosis in dairy cattle. The method was used by conducting tests of RBT, CFT, I-ELISA and commercial I-ELISA to test brucellosis. The test results were calculated sensitivity and specificity, as well as analyzed by calculating the kappa value. The method was used by conducting tests of RBT, CFT, I-ELISA and commercial I-ELISA to test brucellosis. The test results were calculated for sensitivity and specificity, as well as analyzed by calculating the Kappa statistical value. The results of the sensitivity and specificity calculation showed that the indirect enzyme linked immunosorbent assay (I-ELISA) test developed a higher sensitivity (100%) compared to RBT test (93.75%) and commercial I-ELISA (93.75%). The developed I-ELISA specificity (74.68%) was still lower than RBT (89.87%), but higher than commercial I-ELISA (70.52%). The calculation of the statistical value of kappa RBT with CFT showed the kappa value 0.7120 which meaned it had a good agreement, commercial I-ELISA with CFT showed kappa value 0.6165 which meaned it had good suitability, whereas I-ELISA developed with CFT showed kappa value 0.4984 which meaned having a moderate agreement.In conclusion, the indirect enzyme linked immunosorbent assay (I-ELISA) which had been developed had low specificity, but the sensitivity was the highest compared to the commercial I-ELISA test and RBT, so this test was appropriate to be used as a screening test, especially in dairy cows movement into brucellosis-free areas or regions.


1998 ◽  
Vol 5 (5) ◽  
pp. 717-720 ◽  
Author(s):  
E. E. Zijlstra ◽  
N. S. Daifalla ◽  
P. A. Kager ◽  
E. A. G. Khalil ◽  
A. M. El-Hassan ◽  
...  

ABSTRACT The rK39 enzyme-linked immunosorbent assay (ELISA) was compared with the direct agglutination test (DAT) for Leishmania donovani infection in the Sudan. rK39 ELISA proved more sensitive than DAT in diagnosis of kala-azar (93 and 80%, respectively); both tests may remain positive up to 24 months after treatment. For patients with post-kala-azar dermal leishmaniasis and individuals with subclinical infection, rK39 ELISA performed as well as DAT but could detect infection 6 months earlier in ∼40% of patients. Conversion in DAT and rK39 ELISA also occurred in leishmanin skin test (LST)-positive individuals, suggesting active parasite replication (rK39 is an amastigote antigen) in these presumably immune individuals. In contrast to DAT, rK39 ELISA also detected infection in randomly selected LST-positive individuals (in four of six) and endemicity (LST-negative) controls (in one of five). rK39 ELISA appears more sensitive than DAT and may prove an important tool in epidemiological studies.


2005 ◽  
Vol 12 (12) ◽  
pp. 1410-1415 ◽  
Author(s):  
K. M. Kurkjian ◽  
L. E. Vaz ◽  
R. Haque ◽  
C. Cetre-Sossah ◽  
S. Akhter ◽  
...  

ABSTRACT Several serology-based immunoassays are used to diagnose visceral leishmaniasis (VL), a chronic protozoan parasitic disease caused by the Leishmania donovani complex. These tests are primarily designed to diagnose the most severe clinical form of VL, known as kala-azar. However, leishmanial infection is frequently asymptomatic and may manifest only as a positive serologic response or positive leishmanin skin test. We modified a previously described enzyme-linked immunosorbent assay (ELISA) that detects patient antibodies reactive with the recombinant Leishmania protein K39 (rK39) to confirm suspected kala-azar and to detect asymptomatic infection in a community study in Bangladesh. With the inclusion of a standard curve on each ELISA plate, the rK39 ELISA was more repeatable (kappa coefficient of agreement = 0.970) and more reliable compared to the original method (kappa = 0.587, P < 0.001). The cutoff point for a positive antibody response was chosen based on the 99th percentile of the ELISA distribution for the negative-control sera. However, we found that sera from all patients with active kala-azar yielded values more than twice the magnitude of this cutoff. Using receiver-operator characteristic curves, we determined a second cutoff value predictive of kala-azar. Using these criteria, the sensitivity and specificity of the modified ELISA for kala-azar were 97.0% and 98.9%, respectively, for sera from our study population. We hypothesize that individuals with antibody levels greater than the 99th percentile of the negative controls but less than the cutoff point for kala-azar have asymptomatic leishmanial infections.


1978 ◽  
Vol 72 (3) ◽  
pp. 243-253 ◽  
Author(s):  
M. McLaren ◽  
C. C. Draper ◽  
J. M. Roberts ◽  
E. Minter-Goedbloed ◽  
G. S. Ligthart ◽  
...  

2015 ◽  
Vol 8 (1) ◽  
Author(s):  
Sophia Arlena Ulrich ◽  
Kristina Lehnert ◽  
Ursula Siebert ◽  
Christina Strube

1996 ◽  
Vol 59 (1) ◽  
pp. 6-10 ◽  
Author(s):  
PAUL C. BARTLETT ◽  
RONALD J. ERSKINE ◽  
PATRICK GASTON ◽  
PHILIP M. SEARS ◽  
HENDRICUS WILHELMUS HOUDIJK

Recent reports have indicated that the relative sensitivity and specificity of the ELISA test for detection of intramammary infection of cows with Staphylococcus aureus is not as high as originally reported. It has been suggested that antibodies measured by enzyme-linked immunosorbent assay (ELISA) more closely reflect previous infection status rather than current infection status, and that the delay in antibody formation following infection and the persistence of antibodies after elimination of infection may be responsible for some of the discrepancy observed between ELISA and bacterial culture results conducted on the same milk sample. This study (n = 209 cows) was undertaken to determine if an ELISA for S. aureus intramammary infection more closely reflects previous infection status than it does current infection status, and to ascertain whether correction of this time-delay factor substantially improves calculated values of ELISA relative sensitivity and specificity. Receiver-operator curves were constructed to compare different time-related definitions of microbiologic culture results used for comparison with ELISA results. A greater degree of curvature in receiver-operator curves indicated that ELISA results did more closely reflect culture results performed on milk samples taken 1 and 3 weeks previously. Insignificant improvement in sensitivity and specificity occurred when the database was limited to cows (n = 140) with milk production greater than 13.6 kg/day. However, values of sensitivity were all less than or equal to 90%, and values of specificity were all less than 54%.


1991 ◽  
Vol 5 (4) ◽  
pp. 299-301 ◽  
Author(s):  
Krishna Mukerji ◽  
Abhijit Pal ◽  
Kshudiram Naskar ◽  
Dilip K. Ghosh ◽  
Debashish Basu ◽  
...  

2001 ◽  
Vol 8 (2) ◽  
pp. 314-319 ◽  
Author(s):  
Mette Aagaard Strid ◽  
Jørgen Engberg ◽  
Lena Brandt Larsen ◽  
Kamilla Begtrup ◽  
Kåre Mølbak ◽  
...  

ABSTRACT An enzyme-linked immunosorbent assay (ELISA) was adapted to measure immunoglobulin G (IgG), IgM, and IgA classes of human serum antibody toCampylobacter jejuni and Campylobacter coli. Heat-stable antigen, a combination of C. jejuni serotype O:1,44 and O:53 in the ratio 1:1, was used as a coating antigen in the ELISA test. A total of 631 sera from 210 patients with verifiedCampylobacter enteritis were examined at various intervals after infection, and a control group of 164 sera were tested to determine the cut-off for negative results. With a 90th percentile of specificity, IgG, IgM, and IgA showed a sensitivity of 71, 60, and 80%, respectively. By combining all three antibody classes, the sensitivity was 92% within 35 days after infection, whereas within 90 days after infection, a combined sensitivity of 90% was found (IgG 68%, IgM 52%, and IgA 76%). At follow-up of the patients, IgG antibodies were elevated 4.5 months after infection but exhibited a large degree of variation in antibody decay profiles. IgA and IgM antibodies were elevated during the acute phase of infection (up to 2 months from onset of infection). The antibody response did not depend on Campylobacter species or C. jejuniserotype, with the important exception of response to C. jejuni O:19, the serotype most frequently associated with Guillain-Barré syndrome. All of the patients infected with this serotype had higher levels of both IgM (P = 0.006) and IgA (P = 0.06) compared with other C. jejuni and C. coli serotypes.


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