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.

This paper reports attempts to develop a sensitive and inexpensive procedure for rapid diagnosis of cutaneous leishmaniasis at the species level using skin scrapings from patients. The presence of 3 species (Leishmania major, L. tropica and L. infantum) in Israel and the West Bank demonstrates the need for a species-specific detection method in this region. The primer pair Uni21/Lmj4 was developed on the basis of an L. major minicircle sequence but it also amplified other 'Old World' species of Leishmania. Due to species-specific differences in the size of minicircles, these primers can be used in the polymerase chain reaction to answer diagnostic and epidemiological questions.

The most frequent form of cutaneous leishmaniasis (CL) in Israel and the neighbouring territories is due to Leishmania major, which is endemic mainly in the Jordan Valley and in the Rift Valley. CL due to L. tropica is much less common, and in the past only sporadic cases have been reported. In this study we present data obtained during the years 1988-1992 regarding CL in the area. Our clinic has diagnosed a total of 371 leishmaniasis cases, most of whom acquired the infection in the Jordan Valley, mainly during June and July. About one-third of the patients had single lesions, and one-third more than 5 lesions. We also describe an outbreak of leishmaniasis in Kfar Adumim, a village 15 km east of Jerusalem, where leishmaniasis was previously unknown. Parasites were characterized by the polymerase chain reaction and by immunostaining, and found to be both L. tropica and L. major. The localization of the homes of the affected people on a slope where hyraxes were abundant suggests that these animals might have been involved in the transmission of L. tropica in this area.

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These results suggest that topical paromomycin treatment delays the development of cell-mediated immunity but does not affect the levels of immunity that are eventually achieved.