title: Methods incorporating a polymerase chain reaction and restriction fragment length polymorphism and their use as a 'gold standard' in diagnosing Old World cutaneous leishmaniasis.
abstract: Three polymerase chain reaction (PCR) assays - kinetoplast DNA (kDNA) PCR, 7SL PCR, ITS1 PCR - were compared for their ability to detect leishmanial parasites in the skin tissue aspirates of 212 Palestinian suspect cases of cutaneous leishmaniasis (CL). The kDNA PCR was the most sensitive, detecting 156/170 (91.8%) of positive samples confirmed by a set 'gold standard' but of poor specificity in identifying the species of Leishmania. The 7SL PCR detected 154/170 (90.5%) and the ITS1 PCR only 108/170 (63.5%) of the true positives, and both were 100% specific. The highest Kappa coefficient agreement (95% CI) was obtained with the 7SL PCR (0.792 ± 0.098). Restriction analysis of the products from the ITS1 PCR and 7SL PCR enabled identification of species of Leishmania. L. tropica was the predominant cause of CL, with L. major being less frequent.
authors.affiliation: Al-Quds Nutrition and Health Research Institute, Faculty of Medicine, Al-Quds University, Abu-Deis, P.O. Box 20760, West Bank, Palestinian Authority.
authors.forename: Kifaya
authors.initials: K
authors.lastname: Azmi
authors.forename: Abedelmajeed
authors.initials: A
authors.lastname: Nasereddin
authors.forename: Suheir
authors.initials: S
authors.lastname: Ereqat
authors.forename: Lionel
authors.initials: L
authors.lastname: Schnur
authors.forename: Gabriele
authors.initials: G
authors.lastname: Schonian
authors.forename: Ziad
authors.initials: Z
authors.lastname: Abdeen
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D002648:Child; D002675:Child, Preschool; D018105:DNA, Kinetoplast / Q000235:genetics; D005838:Genotype; D006801:Humans; D007891:Leishmania / Q000235:genetics / Q000302:isolation & purification*; D016773:Leishmaniasis, Cutaneous / Q000175:diagnosis* / Q000469:parasitology; D008875:Middle Aged; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D016053:RNA, Protozoan / Q000235:genetics; D012680:Sensitivity and Specificity; D012867:Skin / Q000469:parasitology; D055815:Young Adult
chemical_list: D018105:DNA, Kinetoplast; D016053:RNA, Protozoan