The ability of T2/B4 primers to detect Leishmania infantum among peripheral blood of visceral leishmaniasis patients in Iran

Abstract

Leishmaniasis, caused by protozoa of the genus leishmania, is a zoonotic and anthroponotic disease that is endemic through the tropical and subtropical regions. Twelve million people are affected worldwide and 350 million are at risk. Visceral leishmaniasis in Iran is sporadic in almost all part of Iran and the endemic regions have increased in many districts throughout Iran, the Kalybar, Ahar and Meshkin-shahr districts in East Azarbaijan and Ardabil province in the Northwest of the country. Bone marrow aspiration or biopsy followed by demonstration of leishmania parasites by microscopic and/or cultural examination is the most common diagnosis procedure. These methods are invasive and high risk. The polymerase chain reaction (PCR) has been applied as an analytical method to reveal the presence of small numbers of parasites directly in clinical samples. A PCR-based protocol for the detection of Leishmania infantum parasites in blood was developed and tested with human samples taken from twenty three new visceral leishmaniasis cases referred from endemic area hospitals of Northwest Iran to Pediatric educational hospital of Tabriz University of Medical Sciences. Four different primer pairs were used which targeted genomic and kinetoplast DNAs. The results showed that the PCR assay's sensitivity was significantly dependent on the PCR primers used.