G. Senaldi et al., SEROLOGICAL DIAGNOSIS OF VISCERAL LEISHMANIASIS BY A DOT-ENZYME IMMUNOASSAY FOR THE DETECTION OF A LEISHMANIA DONOVANI-RELATED CIRCULATING ANTIGEN, Journal of immunological methods, 193(1), 1996, pp. 9-15
Field medicine in tropical areas needs laboratory assays which are ine
xpensive and easy to perform. To meet this need a semi-quantitative do
t-enzyme immunoassay (EIA) was developed for the detection of an L. do
novani-related circulating antigen and tested for clinical relevance i
n the diagnosis of visceral leishmaniasis (VL). The dot-EIA probes ser
um spotted on nitrocellulose for the presence of the antigen using a m
onoclonal antibody raised against L. donovani promastigotes, a peroxid
ase-conjugated rabbit anti-mouse immunoglobulin antiserum and chlorona
phthol as peroxidase substrate. The intensity of dot staining by chlor
onaphthol is read by eye and scored, The dot-EIA was used to test the
following groups: 69 patients with VL from Brazil, Kenya, China and Fr
ance, nine patients with cutaneous leishmaniasis, 38 patients with tro
pical diseases other than VL, five patients with rheumatoid arthritis
and 40 health blood donors. The specificity of the assay was 96.7% (3/
92 false positive) and the sensitivity 98.5% (1/69 false negative). A
quantitative EIA for the detection of serum antibodies which makes use
of a crude, soluble L. infantum promastigote extract as capture antig
en and which was used as the reference method, proved to be more speci
fic (98.9%) but similarly sensitive (98.5%), It should be possible to
produce a kit, suitable for large scale application at low cost in ord
er to facilitate routine use of the dot-EIA in the diagnosis of VL.