Identification of Leishmania spp. by Molecular Amp... [J Clin Microbiol. 2011] - PubMed result

Identification of Leishmania spp.by Molecular Amp...[J Clin Microbiol.2011] - PubMed result: "J Clin Microbiol.2011 Jul 13.[Epub ahead of print]
Identification of Leishmania spp.by Molecular Amplification and DNA Sequencing Analysis of a Fragment of the rRNA Internal Transcribed Spacer 2 (ITS2).
de Almeida ME, Steurer FJ, Koru O, Herwaldt BL, Pieniazek NJ, da Silva AJ.
Source

Division of Parasitic Diseases and Malaria, Center for Global Health, Centers for Disease Control and Prevention, Public Health Service, U.S.Department of Health and Human Services, Atlanta, Georgia, United States of America.

Abstract

Isoenzyme analysis of cultured parasites is the conventional approach for Leishmania species identification.Molecular approaches have the potential to be more sensitive and rapid.We designed polymerase chain reaction (PCR) generic primers to amplify a segment of the rRNA Internal Transcribed Spacer 2 (ITS2) from multiple Leishmania species.To validate the selected ITS2 fragment, we tested clinical specimens and compared the species results obtained by the molecular approach (PCR, followed by DNA sequencing analysis) with those from the parasitologic approach (in vitro culture, followed by isoenzyme analysis).Among the 159 patients with clinical specimens positive by both approaches, a total of 8 Leishmania species were identified.The species results were concordant for all but two patients: for one patient, the results were L.(Viannia) guyanensis by the molecular approach versus L.(V.) braziliensis by the parasitologic approach; for the other patient, the results were L.(Leishmania) tropica versus L.(L.) major, respectively.ITS2 PCR, followed by sequencing analysis, can be used to detect and discriminate among Leishmania species.The results confirmed our hypothesis that a region of the ITS2 gene can complement the characterization of Leishmania parasites at the species level.The approach we developed can be used as a diagnostic tool in reference laboratories with adequate infrastructure to perform molecular characterization of pathogens.

PMID:
21752983
[PubMed - as supplied by publisher]

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