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Journal of Clinical Microbiology, April 2003, p. 1419-1422, Vol. 41, No. 4
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.4.1419-1422.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Prospective Value of PCR Amplification and Sequencing for Diagnosis and Typing of Old World Leishmania Infections in an Area of Nonendemicity

Jean-Pierre Gangneux,^1,2* Jean Menotti,¹ Frédéric Lorenzo,¹ Claudine Sarfati,¹ Hélène Blanche,³ Hung Bui,³ Francine Pratlong,⁴ Yves-Jean-François Garin,¹ and Francis Derouin¹

Laboratoire de Parasitologie-Mycologie, Centre Hospitalier Universitaire Lariboisière-Saint-Louis, 75010 Paris,¹ Laboratoire de Parasitologie-Mycologie, Centre Hospitalier Universitaire de Rennes, 35000 Rennes,² Centre d'Etude du Polymorphisme Humain, Hôpital Saint-Louis, 75475 Paris,³ Laboratoire d'Ecologie Médicale et Pathologie Parasitaire, 34090 Montpellier, France⁴

Received 23 September 2002/ Returned for modification 5 November 2002/ Accepted 7 January 2003

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.

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* Corresponding author. Mailing address: Laboratoire de Parasitologie-Mycologie, Rue Henri le Guilloux, 35000 Rennes, France. Phone: 33 2 23 23 44 90. Fax: 33 2 23 23 46 29. E-mail: jean-pierre.gangneux{at}univ-rennes1.fr.

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Journal of Clinical Microbiology, April 2003, p. 1419-1422, Vol. 41, No. 4
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.4.1419-1422.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

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