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 bone 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 vitro cultivation. The PCR was optimized for routine diagnosis: processing of samples with lysis of erythrocytes without isolation of leukocytes, enzymatic prevention of contamination, internal control of the reaction, and ELISA testing in a microtitration plate hybridization. Of 79 samples (33 BMA and 46 BS) from 77 patients without VL, all the results were negative. Fifty-three samples (9 BMA and 44 BS) were obtained from 13 patients with VL: 6 samples drawn during anti-Leishmania treatment were negative whatever the technique used, and 47 samples (9 BMA and 38 BS) were positive with at least one technique. The sensitivities 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 reliable for diagnosing VL in human immunodeficiency virus-infected patients, and blood sampling should be sufficient for the follow-up.
Costa
J M
JM
Laboratoire de Biologie Moléculaire Marcel Dassault, Hôpital américain de Paris, Neuilly sur Seine, France.
Durand
R
R
Deniau
M
M
Rivollet
D
D
Izri
M
M
Houin
R
R
Vidaud
M
M
Bretagne
S
S
D017088:AIDS-Related Opportunistic Infections / Q000150:complications / Q000175:diagnosis* / Q000469:parasitology; D000818:Animals; D001483:Base Sequence; D001853:Bone Marrow / Q000469:parasitology; D017931:DNA Primers / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods* / Q000706:statistics & numerical data; D005069:Evaluation Studies as Topic; D006801:Humans; D007893:Leishmania donovani / Q000235:genetics / Q000302:isolation & purification; D007898:Leishmaniasis, Visceral / Q000150:complications / Q000175:diagnosis* / Q000469:parasitology; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000706:statistics & numerical data; D016053:RNA, Protozoan / Q000097:blood / Q000235:genetics; D012335:RNA, Ribosomal / Q000097:blood / Q000235:genetics; D012680:Sensitivity and Specificity; D013045:Species Specificity; D014776:Virus Cultivation
D017931:DNA Primers; D016053:RNA, Protozoan; D012335:RNA, Ribosomal