LD bodies are identified by light microscopy of the specimen obtained after staining with Giemsa or Leishman stain. The sensitivity of splenic aspirate smear is more than 95% and is regarded as the gold standard for the diagnosis of Kala-azar. 1-3 But the sensitivity of bone marrow smear is only about 60-85% and the procedure is painful whereas splenic aspiration carries the risk of severe hemorrhage. Eventhough this fatal complication is rare in experienced hands, splenic puncture should be

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... ided in patients with a platelet count of less than 40,000/µL and a prothrombin time of more than five seconds over the control. Leishmania can be grown as promastigotes in artificial culture medium, Novy-McNeal Nicolle Medium (NNN), M199 or Grace's medium with 20% fetal calf serum. The parasite can also be demonstrated after inoculating in laboratory animals such as hamster, mice or guinea pig. 1