Diagnosis of American cutaneous leishmaniasis by enzyme immunoassay using membrane antigens of Leishmania (Viannia) braziliensis

Abstract

This study evaluated the reactivity of membrane antigens of Leishmania (Viannia) braziliensis for the diagnosis of ACL by enzyme immunoassay (EIA). Promastigotes of L. (V.) braziliensis were grown in medium 199 and lysed in a sonicator. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting showed that specific proteins of L. (V.) braziliensis (apparent molecular weights 36 kDa and 48-56 kDa) were recognized by sera from ACL patients. These proteins were eluted from the SDS-PAGE and tested in EIA-IgG with sera from ACL patients, healthy individuals, patients with toxoplasmosis, paracoccidioidomycosis, syphilis, tuberculosis, leprosy, and Chagas disease. The EIA-IgG with membrane antigens allowed us to distinguish patients with ACL from healthy individuals and patients with other diseases (P < 0.0001), and showed a sensitivity of 93.3% and specificity of 90.8%, not including Chagas disease patients. 2D-SDS-PAGE followed by Western blotting was performed to improve the characterization of the antigens, and showed a component with isoelectric points near the acid pH side and apparent molecular weights of 48-56 kDa. The results showed good sensitivity and specificity of EIA-IgG with membrane antigens, indicating their potential use for diagnosis of ACL, as well as seroepidemiological surveys and follow-up of clinically cured patients.

Diagnosing Cutaneous leishmaniasis using Fluorescencein SituHybridization: the Sri Lankan Perspective

“…Indirect Immunofluorescence Assay (IFA), Enzyme Linked Immunosorbent Assay (ELISA), Enzyme Immune Assay (EIA), Flow Cytometry and Immunoblotting (Western blot) are the current methods available for serological diagnosis of CL [64]. Some studies have shown Western blot as the most sensitive (100%) [65].…”

“…Some studies have shown Western blot as the most sensitive (100%) [65]. EIAs have a reported sensitivity of 93.3% and a specificity of 90.8% [64].However, these tests are expensive, require laboratory infra-structure and are time consuming. Flow cytometry has been shown to be more sensitive than ELISA but less specific [66].…”

Accuracy of diagnostic tests for American tegumentary leishmaniasis: a systematic literature review with meta‐analyses

“…The opposite occurs with tests that aim to visualise the parasite, which present high specificity and low sensitivity. Molecular diagnoses have also been used, although their utilisation is limited by the need for specialised centres to perform the technique, as well as by their high cost [8][9][10]. Figure 1 describes the clinical path of this review.…”