1921
Volume 78, Issue 1
  • ISSN: 0002-9637
  • E-ISSN: 1476-1645

Abstract

A real-time polymerase chain reaction (PCR) test was developed on the basis of the glucose-6-phosphate dehydrogenase locus that enables identification and quantification of parasites. Using two independent pairs of primers in SYBR-Green assays, the test identified etiologic agents of cutaneous leishmaniasis belonging to both subgenera, () and () in the Americas. Furthermore, use of TaqMan probes enables distinction between () or () from the other () species. All assays were negative with DNA of related trypanosomatids, humans, and mice. The parasite burden was estimated by normalizing the number of organisms per total amount of DNA in the sample or per host glyceraldehyde-3-phosphate dehydrogenase copies. The real-time PCR assay for () subgenus showed a good linear correlation with quantification on the basis of a limiting dilution assay in experimentally infected mice. The test successfully identifies and quantifies in human biopsy specimens and represents a new tool to study leishmaniasis.

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  • Received : 21 Jun 2007
  • Accepted : 17 Oct 2007

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