Molecular diagnosis of visceral leishmaniasis from blood samples using different genetic markers: A simple, sensitive and less invasive diagnostic approach.

Visceral leishmaniasis (VL), or kala-azar, is a deadly disease with high fatality rates if diagnosis and treatment are delayed. Diagnosis is often delayed due to symptoms that mimic other conditions. Sample isolation and diagnostic procedures are labor-intensive and time-consuming. Rapid immunochromatographic tests cannot differentiate active cases from past infections. In the present study we investigated the utility of peripheral blood samples for molecular diagnosis of VL. Whole genomic DNA from the erythrocyte fraction of blood from VL and cutaneous leishmaniasis (CL) suspected patients was used for PCR using multiple markers (k-DNA, ITS-Ⅰ, and 18s rRNA). PCR amplification of k-DNA, ITS-Ⅰ, and 18s rRNA genes yielded positive results in VL symptomatic patients. However, the same PCR approach with peripheral blood samples from CL patients was not significant. Hence, peripheral blood samples can effectively distinguish active VL cases through PCR using multiple markers, offering a less invasive and labor-intensive diagnostic alternative.