Development of a PCR-based method for diagnosis of Leishmania in blood samples

Visceral Leishmaniasis (VL) cases reported in Mediterranean countries and in Northern Europe are becoming increasingly frequent. The past few years several studies have shown Polymerase Chain Reaction to be more effective than the classical methods for the diagnosis of VL in clinical samples. The purpose of this study was the development of a simple, specific and sensitive PCR-based assay for the detection of Leishmania in blood samples. A specific pair of oligonucleotides was designed using conserved sequences of the ssu-rRNA Leishmania infantum gene. Of the 53 blood samples of patients suspected for leishmaniasis that were processed with the newly designed oligonucleotides, 13 were successfully diagnosed positive. The results were confirmed with sequencing and restriction fragment length polymorphism. The lower detection limit of the reported assay was 10 parasites per ml in all seeded samples tested and considered highly satisfactory for diagnosis of Leishmaniasis in blood samples.