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Am. J. Trop. Med. Hyg., 77(1), 2007, pp. 67-72
Copyright © 2007 by The American Society of Tropical Medicine and Hygiene

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Discrimination of Seven Anopheles Species from San Pedro de Urabá, Antioquia, Colombia, by Polymerase Chain Reaction–Restriction Fragment Length Polymorphism Analysis of Its Sequences

Mario A. Zapata, Astrid V. Cienfuegos, Oscar I. Quirós, Martha L. Quiñones, Shirley Luckhart, AND Margarita M. Correa*
Grupo de Microbiología Molecular, Escuela de Microbiología, and Programa para el Estudio y Control de Enfermedades Tropicales, Facultad de Medicina, Universidad de Antioquia, Medellín, Colombia; Department of Medical Microbiology and Immnunology, School of Medicine, University of California, Davis, California

Accurate identification of anopheline species is essential for vector incrimination and implementation of appropriate control strategies. Several anopheline species are considered important malaria vectors in Colombia; however, species determination is complicated by cryptic morphology and intra-individual variation. We describe polymerase chain reaction–restriction fragment length polymorphism (PCR-RFLP) of internal transcribed spacer 2 (ITS2) sequences for differentiation of seven Anopheles species collected in a locality in Antioquia, Colombia, with high levels of malaria transmission. Each of these seven species can be identified by unique AluI PCR-RFLP restriction patterns. Comparisons of morphologic identification with molecular identification of voucher specimens confirmed species designation for 886 wild-caught anophelines. This new method can be used as a diagnostic tool for discrimination of anopheline species of medical importance in this region, some of which have overlapping morphologic characters and for conducting complementary studies where rapid and accurate identification of large numbers of specimens is needed.

Received December 18, 2006. Accepted for publication March 14, 2007.

Acknowledgments: We are grateful to Dr. Jan E. Conn (Wadsworth Center, New York State Department of Health, Albany, NY) for editorial comments. We also thank Dr. Armando Galeano M. (coordinator) and personnel of the Vector Control Group, Dirección Seccional de Salud de Antioguia (DSSA); Dr. Iván D. Vélez (director), Freddy Ruiz, and personnel at Programa para el Estudio y Control de Enfermedades Tropicales, Universidad de Antioquia; Juan Santiago Zuluaga (Corporación para Investigaciones Biológicas) and staff at the field collecting sites for their support; and Dr. Richard Wilkerson (Walter Reed Smithsonian Institute, Suitland, MD) for assistance.

Financial support: This study was supported by the Gorgas Memorial Institute of the American Society of Tropical Medicine and Hygiene, Comité para el Desarrollo de la Investigación, Universidad de Antioquia, and Fundación para la Promoción de la Investigación y la Tecnología del Banco de la Republica, Colombia.

* Address correspondence to Margarita M. Correa, Grupo de Microbiología Molecular, Escuela de Microbiología, Universidad de Antioquia, Calle 67 No. 53-108, Bloque 5-430, Medellín, Colombia. E-mail: mcorrea{at}quimbaya.udea.edu.co

Authors’ addresses: Mario A. Zapata, Astrid V. Cienfuegos, Oscar I. Quirós, and Margarita M. Correa, Grupo de Microbiología Molecular, Escuela de Microbiología, Universidad de Antioquia, Calle 67 No. 53-108, Bloque 5-430, Medellín, Colombia, Telephone: 57-4-210-5494, Fax: 57-4-210-5498, E-mail: mcorrea{at}quimbaya.udea.edu.co. Martha L. Quiñones, Departamento de Salud Pública, Facultad de Medicina, Universidad Nacional de Colombia, Carrera 30 Calle 45, Bogotá DC, Colombia, Telephone: 57-1-316-5000 extension 15078, Fax: 57-1-316-5175. Shirley Luckhart, Department of Medical Microbiology and Immunology, School of Medicine, University of California at Davis, 3437 Tupper Hall, 1 Shields Avenue, Davis, CA 95616.

Reprint requests: Margarita M. Correa, Grupo de Microbiología Molecular, Escuela de Microbiología, Universidad de Antioquia, Calle 67 No. 53-108, Bloque 5-430, Medellín, Colombia.




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Improved Molecular Technique for the Differentiation of Neotropical Anopheline Species
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