title: Prospective value of PCR amplification and sequencing for diagnosis and typing of old world Leishmania infections in an area of nonendemicity.
abstract: We assessed the prospective value of PCR amplification of a repetitive sequence from Leishmania nuclear DNA and sequencing for the diagnosis and typing of Old World Leishmania infection in an area of nonendemicity. During this 42-month study, 29 of 168 consecutive samples were examined and classified as positive for Leishmania by direct examination and/or in vitro culture. This molecular approach showed excellent sensitivity (97%) and specificity (100%) compared to direct examination (86 and 100%, respectively) and in vitro culture (72 and 100%, respectively). Isoenzymatic and molecular typing allowed similar identification for 12 samples. Besides, PCR and subsequent sequencing of DNA products permitted the species identification of 14 samples for which parasite culture remained negative or did not allow isoenzymatic characterization, indicating the complementarity of parasitological and molecular tools.
authors.affiliation: Laboratoire de Parasitologie-Mycologie, Centre Hospitalier Universitaire Lariboisière-Saint-Louis, 75010 Paris, France. jean-pierre.gangneux@univ-rennes1.fr
authors.forename: Jean-Pierre
authors.initials: JP
authors.lastname: Gangneux
authors.forename: Jean
authors.initials: J
authors.lastname: Menotti
authors.forename: Frédéric
authors.initials: F
authors.lastname: Lorenzo
authors.forename: Claudine
authors.initials: C
authors.lastname: Sarfati
authors.forename: Hélène
authors.initials: H
authors.lastname: Blanche
authors.forename: Hung
authors.initials: H
authors.lastname: Bui
authors.forename: Francine
authors.initials: F
authors.lastname: Pratlong
authors.forename: Yves-Jean-François
authors.initials: YJ
authors.lastname: Garin
authors.forename: Francis
authors.initials: F
authors.lastname: Derouin
mesh_terms: D000818:Animals; D001483:Base Sequence; D003470:Culture Media; D016054:DNA, Protozoan / Q000032:analysis; D006801:Humans; D007527:Isoenzymes / Q000032:analysis; D007891:Leishmania / Q000145:classification* / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification*; D016773:Leishmaniasis, Cutaneous / Q000175:diagnosis* / Q000469:parasitology; D007898:Leishmaniasis, Visceral / Q000175:diagnosis* / Q000469:parasitology; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D011237:Predictive Value of Tests; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D013045:Species Specificity
chemical_list: D003470:Culture Media; D016054:DNA, Protozoan; D007527:Isoenzymes