Real-Time PCR-based diagnosis of human visceral leishmaniasis using urine samples

Abstract

AbstractBackgroundDiagnosis of visceral leishmaniasis (VL) through the detection of its causative agents namely Leishmania donovani and L. infantum is traditionally based on Giemsa-stained smears of bone marrow, spleen aspirates, liver or lymph node. Collection of these samples involve invasive procedures and carries the risk of fatal hemorrhage especially during splenic aspiration. Earlier, we reported a Polymerase Chain Reaction (PCR)-based diagnosis of L. donovani in peripheral blood using a novel set of PCR primers with absolute specificity (Khatun et al. 2017). Using the same set of primers and PCR conditions, here we describe diagnosis of L. donovani from urine, for a non-invasive, rapid and safe diagnosis.MethodsDiagnosis of L. donovani was carried out using urine samples collected from clinically diagnosed VL patients (n=23) of Bangladesh in Real Time PCR. Test results were validated by comparing blood samples from the same set of patients. Sensitivity and specificity of this diagnosis was analyzed using retrospective bone marrow samples, collected earlier from confirmed VL patients (n=19) (Khatun et al. 2017).ResultsThe method showed 100% sensitivity in detecting VL in urine and corresponding blood samples and bone marrow samples, as well as 100% specificity in control groups.ConclusionUrine-based diagnosis could be a patient-friendly, non-invasive approach for VL detection with precision and perfection.