|dc.contributor.author||Schnur, Lionel F.||
|dc.identifier.citation||Azmi K, Schonian G, Schnur LF, Nasereddin A, Ereqat S, et al. (2013) Development of Assays Using Hexokinase and Phosphoglucomutase Gene Sequences That Distinguish Strains of Leishmania tropica from Different Zymodemes and Microsatellite Clusters and Their Application to Palestinian Foci of Cutaneous Leishmaniasis. PLoS Negl Trop Dis 7(9): e2464. doi:10.1371/journal.pntd.0002464||en_US
|dc.description.abstract||Background/Objectives: Palestinian strains of L.tropica characterized by multilocus enzyme electrophoresis (MLEE) fall into
two zymodemes, either MON-137 or MON-307.
Methodology/Principle Findings: Assays employing PCR and subsequent RFLP were applied to sequences found in the
Hexokinase (HK) gene, an enzyme that is not used in MLEE, and the Phosphoglucomutase (PGM) gene, an enzyme that is
used for MLEE, to see if they would facilitate consigning local strains of L.tropica to either zymodeme MON-137 or
zymodeme MON-307. Following amplification and subsequent double digestion with the restriction endonucleases MboI
and HaeIII, variation in the restriction patterns of the sequence from the HK gene distinguished strains of L.tropica, L.major
and L.infantum and also exposed two genotypes (G) among the strains of L.tropica: HK-LtG1, associated with strains of
L.tropica of the zymodemes MON-137 and MON-265, and HK-LtG2, associated with strains of L.tropica of the zymodemes
MON-307, MON-288, MON-275 and MON-54. Following amplification and subsequent digestion by the restriction
endonuclease MboI, variation in the sequence from the PGM gene also exposed two genotypes among the strains of
L.tropica: PGM-G1, associated only with strains of L.tropica of the zymodeme MON-137; and PGM-G2, associated with strains
of L.tropica of the zymodemes MON-265, MON-307, MON-288, MON-275 and MON-54, and, also, with six strains of L.major,
five of L.infantum and one of L.donovani. The use of the HK and PGM gene sequences enabled distinction the L.tropica
strains of the zymodeme MON-137 from those of the zymodeme MON-265. This genotyping system ‘correctly’ identified
reference strains of L.tropica of known zymodemal affiliation and also from clinical samples, with a level of sensitivity down
to ,1 fg in the case of the former and to 1 pg of DNA in the case of the latter.
Conclusions/Significance: Both assays proved useful for identifying leishmanial parasites in clinical samples without
resource to culture and MLEE.||en_US
|dc.description.sponsorship||This study was supported by the Deutsche Forschungsgemeinschaft (DFG) as part of a German-Israeli-Palestinian cooperative project on the Emergence of Cutaneous Leishmaniasis in the Middle East: ‘An investigation of Leishmania tropica in the Palestinian Authority and Israel’ (SCHO 448/8-1). The
funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
We thank Christophe Ravel and Gert de Auwera, at Universite´
Montpellier 1 and Centre Hospitalier Universitaire de Montpellier,
Laboratoire de Parasitologie and Centre National de Reference des
Leishmania, Montpellier, France, for their help for providing DNA from L.
tropica strains of known zymodeme that were used in this study.||en_US
|dc.title||Development of Assays Using Hexokinase and Phosphoglucomutase Gene Sequences That Distinguish Strains of Leishmania tropica from Different Zymodemes and Microsatellite Clusters and Their Application to Palestinian Foci of Cutaneous Leishmaniasis||en_US