Diagnosis of Visceral Leishmaniasis by Enzyme-Linked Immunosorbent Assay Using Urine Samples

Abstract

A diagnostic method has been developed to detect anti-Leishmania donovani immunoglobulin G (IgG) in urine by enzyme-linked immunosorbent assay (ELISA). In measuring anti-L. donovani IgG, IgA, and IgM in urine, the method performed best in the detection of IgG. The sensitivity and specificity of the assay were determined with panels of urine samples from 62 visceral leishmaniasis (VL) patients, 59 healthy controls from areas of endemicity, 53 healthy controls from areas of nonendemicity, 59 malaria patients, 13 tuberculosis patients, 23 cutaneous leishmaniasis patients, and 7 patients with other diseases. Using L. donovani promastigote crude antigen, the test had 93.5% sensitivity (58 positives of 62 VL patient samples) and 89.3% specificity (191 negatives of 214 non-VL patient samples). The ELISA with acetone-treated L. donovani promastigote antigen raised the sensitivity and specificity to 95.0 and 95.3%, respectively. Western blot analysis revealed that most of the samples that cross-reacted with crude antigen in ELISA did not recognize any antigenic component of L. donovani crude antigen. We also checked 40 serum samples from the same group of VL patients for anti-L. donovani IgG and got 90.0% sensitivity with both crude and acetone-treated antigens. As collection of urine is much easier than collection of serum, the detection of anti-L. donovani IgG in urine with acetone-treated antigen will be useful in epidemiological studies. It could be an adjunct of laboratory diagnosis.

Detection of Anti-LeishmaniaImmunoglobulin G Antibodies in Urine Specimens of Dogs with Leishmaniasis

“…Moreover, several infectious diseases are diagnosed by the finding of specific antibodies in urine, such as Helicobacter pylori infection (18), filariasis (17), or schistosomiasis (35). The presence of anti-Leishmania antibodies in the urine of human patients with visceral leishmaniasis (16,20) and in the urine of L. donovani-infected hamsters (34) has also been described. In contrast, there is a lack of information about immunoglobulins in the urine of healthy and sick dogs.…”

“…All positive samples (urine and serum samples) recognized numerous polypeptide fractions of the L. infantum antigen with masses ranging from 12 to 85 kDa, with most bands at 16 to 69 kDa. The most frequent polypeptide fractions observed in both urine and serum samples were those of 14,16,20,30,31,48, and 69 kDa. The immunoblot analysis did not reveal any polypeptide fractions of the L. infantum antigen in ELISA-negative urine samples.…”

Comparison of New Diagnostic Tools for Management of Pediatric Mediterranean Visceral Leishmaniasis

“…Many of these problems may be avoided by searching for antigens in the urine. Several studies have demonstrated Leishmania antigens in the urine of VL patients and animals using different techniques including double countercurrent immunoelectrophoresis (22), ELISA (3,20,36), and Western blotting (13). Recently, a latex agglutination test (KAtex kit) to detect Leishmania antigen in urine showed 100% specificity and 47 to 100% sensitivity in different studies with immunocompetent patients (2,32) and 96 to 100% specificity and 85 to 100% sensitivity in immunodepressed patients (31,37).…”

An enzyme-linked immunosorbent assay (ELISA) for the detection of IgM antibodies against Leishmania chagasi in dogs

“…Several serological tests have been developed and evaluated for the diagnosis of VL including indirect fluorescent antibody test (IFAT); enzyme-linked immunosorbent assay (ELISA); Dot-ELISA; immunoblot analysis; and direct agglutination test (DAT) (Islam et al 2002). The presence of antiparasite antibodies is routinely used as a marker of infection with the production of antibody as the definition of infection or challenge.…”