Comparison of PCR assays for diagnosis of cutaneous leishmaniasis.
Three PCR assays for diagnosing leishmaniasis were compared and validated against parasite cultures and microscopic evaluation of stained tissue smears using 92 specimens from suspected cases of cutaneous leishmaniasis (CL) in Israel and the West Bank. Samples from imported and locally acquired disease were examined. The kinetoplast DNA (kDNA) PCR showed the highest sensitivity (98.7%) of any assay, correctly diagnosing 77/78 of the confirmed positive samples, followed by the rRNA gene internal transcribed spacer 1 (ITS1) PCR (71/78 positive, 91.0% sensitivity) and then the spliced leader mini-exon PCR (42/78 positive, 53.8% sensitivity). Either parasite culture or microscopy alone detected 62.8% (49/78) or 74.4% (58/78) of the positive specimens, respectively, while culture and microscopy together improved overall sensitivity to 83.3% (65/78). Except for the kDNA PCR that had six false positives, all other assays were 100% specific. Further, restriction enzyme analysis of the ITS1 PCR product enabled identification of 74.6% of the positive samples, which included strains of Leishmania major (50.9%), Leishmania tropica (47.2%), and the Leishmania braziliensis complex (1.9%). This suggests that a PCR using kDNA should be used for the diagnosis of CL and that an ITS1 PCR can be reliably used for the diagnosis of CL when rapid species identification is needed.
Bensoussan
Esther
E
Department of Parasitology, P.O. Box 12272, Hebrew University-Hadassah Medical School, Jerusalem 91120, Israel.
Nasereddin
Abedelmajeed
A
Jonas
Flory
F
Schnur
Lionel F
LF
Jaffe
Charles L
CL
D000818:Animals; D018105:DNA, Kinetoplast / Q000235:genetics; D006801:Humans; D007892:Leishmania braziliensis / Q000145:classification / Q000302:isolation & purification*; D007895:Leishmania tropica / Q000235:genetics / Q000302:isolation & purification; D016773:Leishmaniasis, Cutaneous / Q000175:diagnosis* / Q000453:epidemiology / Q000469:parasitology; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D012680:Sensitivity and Specificity
D018105:DNA, Kinetoplast