Development of an Indirect Fluorescent Antibody (Ifa) Assay for the Detection of Leishmania Rna Virus 2 (Lrv2) in Leishmania Parasites Publisher



Hajjaran H¹ ; Ebadizadeh M¹ ; Ataeipirkooh A² ; Mohebali M¹ ; Samimirad K³ ; Saberi R⁴ ; Naddaf SR⁵ Show Affiliations
Source: Iranian Journal of Parasitology Published:2022

Abstract

Background: Detection of Leishmania RNA virus (LRV) in Old World Leishmania species atheir possible role in the disease prognosis requires sensitive and specific methods, preferabindependent of the viral genome. We aimed to develop an indirect immunofluorescence anbody (IFA) assay to detect LRV in the Old World Leishmania parasites. Methods: Clinical samples were collected from 86 cutaneous leishmaniasis (CL) patients in dferent endemic areas of CL in Iran, during 2017-2019. For antibody preparation, the viruswere obtained from sediment of an LRV-infected L. major culture-using freeze and thaw cyclfollowed by gradient cesium chloride centrifugation. The purified viruses were used to immunia male 3-4 months rabbit. Various dilutions of the LRV-immunized rabbit's serum and a conjgated antibody were deployed to detect LRV in 48 isolates by IFA assay. Results: LRV virus was detected in four of the 48 CL cases using IFA method. Amplification a partial fragment of RNA-dependent RNA polymerase (RdRp) gene from the isolates cofirmed the IFA results. In phylogeny, the generated RdRp sequences from four isolates wegrouped with the other Old World LRVs, but separate from L. aethiopica LRVs, which appearas a highly supported distinct clade. Conclusion: Further optimization of this approach to detect the LRV directly in lesion scrapings can make it a more reliable tool for field studies and disclosing the virus's possible role in disseminating and unusual clinical features. © 2022 Hajjaran et al. Published by Tehran University of Medical Sciences.