Visual assessment of parasitic burden in infected macrophage by plasmonic detection of leishmania specific marker RNA

Abstract

Visceral leishmaniasis is a neglected tropical disease and may prove fatal if not diagnosed and treated early. The amastigotes of Leishmania donovani nest in the macrophage of human host and thus, determination of parasitic burden in the infected macrophages has been the most crucial step in diagnosis, dose determination and medical management of relapse cases of this fatal disease. Microscopic count following Giemsa staining and other morphological analysis are the classical ways vastly used in the resource stringent endemic areas. The current method introduced a high throughput, rapid, cheap, non-gel, non-PCR and nonculture based visual detection platform employing salt triggered aggregation of gold nanoparticle in presence of extracted total RNA from infected macrophages and leishmania specific oligo-nucleotide probe to determine the parasite burden in macrophages. Amastigote's small subunit ribosomal RNA (SSU rRNA, PMID 1565128) was used as the leishmania specific marker and its abundance in the total RNA extracts of infected macrophages were determined by this visual colorimetric assay.