Diagnosis of leishmaniasis has always been a major challenge as its clinical features resemble some other commonly occurring diseases such as tuberculosis, typhoid, and malaria. Reliable laboratory methods become important for differential diagnosis. Demonstration of the parasites in stained preparations of bone marrow and splenic aspirates being risky and invasive is still the gold standard for diagnosis. Serological tests utilizing rapid immunochromatographic formats or rK39 in enzyme linked immune sorbent assay, immunoblotting, direct agglutination test have complications related to high proportions of positive asymptomatic individuals and the inability to diagnose a relapse. Among the molecular techniques, polymerase chain reaction is the most commonly used technique that is successfully implied for diagnosis. This review provides updated information on the recent developments in the field of diagnosis in leishmaniasis, various methods utilized with their advantages and limitations.