Appraisal of various random amplified polymorphic DNA-polymerase chain reaction primers for Leishmania identification

Pathogenic Leishmania can cause a variety of symptoms ranging from asymptomatic or mild infections to severe mucocutaneous disease, partly according to the species of Leishmania involved. Genomic and kinetoplast-derived DNA probes as well as species complex-specific kinetoplast-derived polymerase chain reaction (PCR) primers have been successful for parasite identification in epidemiologic and taxonomic studies. However, the lack of a species DNA probe or PCR primer set, their relatively poor availability, and unknown comparative sensitivity and specificity have precluded their routine and widespread use as identification tools in many laboratories. This study addresses this problem for 28 different random amplified polymorphic DNA (RAPD)-PCR primers that have been assessed regarding their sensitivity, specificity, and reliability for distinguishing each of four closely related New World Leishmania species. The degree of relatedness between species was quantified and estimates were made of the accuracy and precision of the determinations. The results compared well with standard methods for Leishmania classification. The application of RAPD-PCR for screening isolates for possible interspecific hybrids is also demonstrated.