Detection of Leishmania donovani in peripheral blood of asymptomatic individuals in contact with patients with visceral leishmaniasis

Abstract

Background: The majority of individuals infected with Leishmania donovani complex remain asymptomatic. They may act as transmission reservoirs for visceral leishmaniasis (VL). We investigated sero-prevalence of L. donovani complex amongst those closely associated with patients with VL and whether these sero-reactive individuals had Leishmania parasites in their peripheral blood. Other risk factors were also investigated.

Methods: A total of 257 individuals in contact with patients with VL were tested for anti-Leishmania antibodies by rK39 immunochromatographic test (rK39 ICT), ELISA using promastigote antigen (p-ELISA) and indirect fluorescent antibody test (IFAT). Buffy coats of rK39 ICT positive individuals were cultured; sero-reactive buffy coats were tested for Leishmania DNA by ITS1 PCR. DNA obtained from culture was sequenced to confirm Leishmania species. Risk factors were evaluated for each sero-positive sample.

Results: The results showed 29.2% (75/257) prevalence by serological tests: 14.4% (37/257) were positive by rK39 ICT, 25.3% (65/257) by p-ELISA, 18.3% (47/257) by IFAT and 10.9% (28/257) by all three serological methods. Ten percent (3/30) of cultures were positive for Leishmania promastigotes. Only 3% (2/74) sero-reactive buffy coats were positive for DNA; sequence analysis revealed L. donovani species. Significant risk factors were age, working as farmers, domestic animals in household and proximity to animal shelters.

Conclusions: Asymptomatic family members of patients with VL can carry live L. donovani in peripheral blood and may act as potential reservoirs.

Genbank accession number: BankIt1863680 Leishmania KT921417 (DNA sequences of the ribosomal ITS1 region of L. donovani).

Keywords: Asymptomatic infection; Bangladesh; Culture; Leishmania; PCR; Serology.