Visceral leishmaniasis (VL) is a parasitic disease and serious public health problem. The most severe form of all leishmaniases, it is fatal if untreated. Currently, it affects close to 540 people per year in Algeria, mainly in Kabylie, where it is fatal in 6% of all cases. This disease primarily affects children. Its diagnosis is based principally on visualization of parasites in bone marrow aspirate. Serologic studies are also used to diagnose VL, as is molecular biology, which has been demonstrated to be both faster and more sensitive. The aim of our study was to evaluate the utility of real-time polymerase chain reaction (RT-PCR) in the diagnosis of VL in endemic areas. Of the 108 samples analyzed, 32 were positive according to RT-PCR (29.6%); microscopy yielded positive result in 24% and serology in 25.9%. RT-PCR increased the overall number of diagnoses detected from 26 to 32, a 20% improvement over microscopic methods. In view of the lethal consequences of failure to diagnose this disease, any improvement in diagnostic methods provides direct medical benefit.