Diagnostic Microbiology & Infectious Disease
Volume 66, Issue 2 , Pages 175-180, February 2010

Leishmania spp. identification by polymerase chain reaction–restriction fragment length polymorphism analysis and its applications in French Guiana

  • Stéphane Simon

      Affiliations

    • Equipe EA 3593, Faculté de Médecine, Université des Antilles et de la Guyane, Cayenne, French Guiana
  • ,
  • Vincent Veron

      Affiliations

    • Equipe EA 3593, Faculté de Médecine, Université des Antilles et de la Guyane, Cayenne, French Guiana
  • ,
  • Bernard Carme

      Affiliations

    • Equipe EA 3593, Faculté de Médecine, Université des Antilles et de la Guyane, Cayenne, French Guiana
    • Laboratoire Hospitalo-Universitaire de Parasitologie-Mycologie (LHUPM), Centre Hospitalier de Cayenne, French Guiana
    • Corresponding Author InformationCorresponding author. Laboratoire Hospitalo-Universitaire de Parasitologie et Mycologie Médicale, Equipe EA 3593, Centre Hospitalier de Cayenne et Faculté de Médecine de l'Université des Antilles et de la Guyane, Rue des Flamboyants, BP 6006, 97336 Cayenne, French Guiana. Tel.: +594-594-39-53-09; fax: +594-594-39-53-09.

Received 9 May 2009; accepted 16 August 2009. published online 27 September 2009.

Abstract 

Leishmania (Viannia) guyanensis was for many years the only species commonly identified in French Guiana, but precise species identifications were quite rare. We describe a new restriction fragment length polymorphism–polymerase chain reaction technique using a 615-bp fragment of the RNA polymerase II gene and 2 restriction enzymes, TspRI and HgaI. Seven reference strains (Leishmania (Leishmania) amazonensis, Leishmania (Viannia) lainsoni, Leishmania (Viannia) braziliensis, L. (V.) guyanensis, Leishmania (Viannia) naiffi, Leishmania (Leishmania) major, Leishmania (Leishmania) infantum) and 112 clinical samples from positive lesions were used for the development of the technique. The rates of positive species identification were 85.7% for punch skin biopsy specimens, 93.1% for positive Giemsa-stained smears, and 100% for positive culture supernatants. In the framework of cutaneous leishmaniasis species surveillance for the 2006 to 2008 period, parasite identification was carried out for 199 samples from different patients. The prevalence of the various Leishmania spp. was 84.4% for L. (V.) guyanensis, 8.0% for L. (V.) braziliensis, 5.0% for L. (L.) amazonensis, and 2.6% for L. (V.) lainsoni. L. (V.) braziliensis seems to be locally an emerging pathogen.

Keywords: Cutaneous leishmaniasis, Leishmania, Diagnosis, Species identification, PCR, RFLP, French Guiana

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 This work was supported by the University of the French West Indies and French Guiana and the French Ministère de l'Enseignement Supérieur et de la Recherche Scientifique.

PII: S0732-8893(09)00356-3

doi:10.1016/j.diagmicrobio.2009.08.013

Diagnostic Microbiology & Infectious Disease
Volume 66, Issue 2 , Pages 175-180, February 2010