1887

Abstract

Following antigen preparation procedures similar to those of the direct agglutination test (DAT), an IgG ELISA employing intact -mercaptoethanol (-ME)-treated promastigotes was developed. The performance of the -ME ELISA thus developed was assessed in patients with confirmed visceral leishmaniasis (VL), revealing slightly lower sensitivity (39/40=97.5 %) than that of the DAT (40/40=100 %). When challenged with sera of individuals with non-VL conditions, including leukaemia and African trypanosomiasis, the specificity of the -ME ELISA was 100 % (158/158), compared to 98.8 % (156/158) for DAT. In an endemic population (=145) manifesting a clinical suspicion of VL, results obtained with the -ME ELISA were highly concordant with those of DAT, both in the seropositive (65/68=95.6 %) and seronegative (77/80=96.3 %) groups. Furthermore, the incorporated intact antigen demonstrated higher sensitivity in ELISA (16/18=88.9 %) than the water-soluble equivalent (13/18=72.2 %). The stability of the formaldehyde-fixed antigen (2 months at 4 °C) in -ME ELISA, as well as the option for direct testing of whole-blood samples and visual reading of results (within 2 h, compared to 18 h for DAT), advocate the simultaneous application of the technique with DAT for confirmation of VL in laboratories with limited facilities.

Loading

Article metrics loading...

/content/journal/jmm/10.1099/jmm.0.46643-0
2006-09-01
2019-12-15
Loading full text...

Full text loading...

/deliver/fulltext/jmm/55/9/1193.html?itemId=/content/journal/jmm/10.1099/jmm.0.46643-0&mimeType=html&fmt=ahah

References

  1. Burns, J. M., Shreffer, W. G., Benso, D. R., Ghalib, H. W., Badaro, R. & Reed, S. G. ( 1993; ). Molecular characterization of a kinesin-related antigen of Leishmania chagasi that detects specific antibody in African and American visceral leishmaniasis. Proc Natl Acad Sci U S A 90, 775–779.[CrossRef]
    [Google Scholar]
  2. Harith, A. E., Kolk, A. H. J., Leewenburg, J., Muigai, R., Jelsma, T. & Kager, P. A. ( 1988; ). Improvement of a direct agglutination test for field studies of visceral leishmaniasis. J Clin Microbiol 26, 1321–1325.
    [Google Scholar]
  3. Harith, A. E., Chowdhury, S., Al Massum, A., Samiao-Santos, S., Karim, E., El Safi, S. & Haque, I. ( 1995; ). Evaluation of cleaving agents other than trypsin in direct agglutination test for further improving diagnosis of visceral leishmaniasis. J Clin Microbiol 33, 1984–1988.
    [Google Scholar]
  4. Hommel, M., Peters, W., Ranque, J., Quilici, M. & Lanotte, G. ( 1978; ). The micro-ELISA technique in the serodiagnosis of visceral leishmaniasis. Ann Trop Med Parasitol 72, 213–218.
    [Google Scholar]
  5. Mohammed, E. A. R., Wright, P. A., Kager, P. A., Laarman, J. J. & Pondman, K. W. ( 1985; ). ELISA using intact promastigotes for immunodiagnosis of kala-azar. Trans R Soc Trop Med Hyg 79, 344–350.[CrossRef]
    [Google Scholar]
  6. Sadigursky, M. & Brodskyn, C. I. ( 1986; ). A new liquid medium without blood or serum for culture of haemoflagellates. Am J Trop Med Hyg 35, 942–944.
    [Google Scholar]
  7. Shiddo, S. A. ( 1995; ). Visceral leishmaniasis in Somalia: immunodiagnostic and epidemiological aspects. PhD thesis, Swedish Institute for Infectious Disease Control.
  8. Voller, A., Bidwell, D. & Barlett, A. ( 1980; ). Enzyme-linked immunosorbent assay. In Manual of Clinical Immunology, pp. 359–371. Edited by N. Rose & H. Friedman. Washington, DC: American Society for Microbiology.
  9. Zijlstra, E. E., Nur, Y., Desjeux, P., Khalil, E. A. G., El-Hassan, A. M. & Groen, J. ( 2001; ). Diagnosing visceral leishmaniasis with the recombinant K39 strip test: experience from Sudan. Trop Med Int Health 6, 108–113.[CrossRef]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jmm/10.1099/jmm.0.46643-0
Loading
/content/journal/jmm/10.1099/jmm.0.46643-0
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error