PCR-ELISA for diagnosis of mucocutaneous leishmaniasis

In this work we demonstrate that the PCR-ELISA technique is sufficiently se nsitive and specific for use as a diagnostic test in cases of mucocutaneous leishmaniasis. DNA was extracted from cultures of Leishmania braziliensis, Leishmania infantum, Leishmania tropica, Leishmania mexicana, Trypanosoma cruzi, and blood samples from individuals who presented a clinical diagnosi s of leishmaniasis as well as from healthy individuals. The DNA was PCR amp lified and the product obtained was hybridised with a biotin-labelled probe , the sequence of which was designed in our laboratory. The result of the h ybridisation was visualised by means of an ELISA technique using antifluore scein antibody labelled with alkaline phosphatase and p-nitrophenylphosphat e (pNFF) as chromogen. The optical density of the products of the pNFF hydr olysis was quantified in a spectrophotometer at a wavelength of 405 nm. Usi ng this technique the percentage of detection was 83.3% in blood samples fr om patients clinically diagnosed as having mucocutaneous leishmaniasis. No false positive results were obtained. (C) 1999 Elsevier Science B.V. All ri ghts reserved.