PCR ENZYME-LINKED-IMMUNOSORBENT-ASSAY FOR DIAGNOSIS OF LEISHMANIASIS IN HUMAN IMMUNODEFICIENCY VIRUS-INFECTED PATIENTS

A PCR enzyme-linked immunosorbent assay (ELISA) involving the use of b one marrow aspirates (BMA) and blood samples (BS) for the diagnosis of visceral leishmaniasis (VL) in human immunodeficiency virus-infected patients was developed with primers selected from the sequence of the small-subunit rRNA gene and compared with direct examination and in vi tro cultivation, The PCR was optimized for routine diagnosis: processi ng of samples with lysis of erythrocytes without isolation of leukocyt es, enzymatic prevention of contamination, internal control of the rea ction, and ELISA testing in a microtitration plate hybridization, Of 7 9 samples (33 BMA and 46 BS) from 77 patients without VL, all the resu lts were negative, Fifty-three samples (9 BMA and 44 BS) were obtained from 13 patients with VL: 6 samples drawn during anti-Leishmania trea tment were negative whatever the technique used, and 47 samples (9 BMA and 38 BS) mere positive with at least one technique, The sensitiviti es were 51% (24 of 47), 81% (38 of 47), and 98% (46 of 47) for direct examination, culture, and PCR, respectively, Thus, PCR ELISA is reliab le for diagnosing VL in human immunodeficiency virus-infected patients , and blood sampling should be sufficient for the follow-up.