AJTMH Transactions of the Royal Society of Tropical Medicine and Hygiene
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Am. J. Trop. Med. Hyg., 80(2), 2009, pp. 236-238
Copyright © 2009 by The American Society of Tropical Medicine and Hygiene

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SHORT REPORT


A Multiplex PCR Assay for Simultaneous Genotyping of kdr and ace-1 Loci in Anopheles gambiae

Athina Kazanidou, Dimitra Nikou, Maria Grigoriou, John Vontas, AND George Skavdis*
Department of Molecular Biology and Genetics, Democritus University of Thrace, Alexandroupolis, Greece; Vector Group, Liverpool School of Tropical Medicine, Liverpool, United Kingdom; Laboratory of Pesticide Science, Agricultural University of Athens, Athens, Greece

 

ABSTRACT

The selection of insecticide-resistant genotypes in Anopheles gambiae, the most important malaria vector in Africa, makes disease control problematic in several endemic areas. The early detection and monitoring of resistance associated mutations in field mosquito populations is essential for the application of successful insecticide-based control interventions. Currently, the surveillance of these mutations is performed using individual assays, some of which require sophisticated and expensive equipment. Here we describe a novel multiplex polymerase chain reaction–based assay for detecting simultaneously the five single nucleotide polymorphisms in the voltage-gated sodium channel and the ace-1 genes, which have been associated with the mosquito response to most commonly used insecticides.

Received August 2, 2008. Accepted for publication November 8, 2008.

Acknowledgments: The authors thank Dr. Fabrice Chandre (Institut de Recherche pour le Developpement/Centre de Recherche Entomologique de Cotonou, Benin) for generously providing the ACER-KIS and ACER-KIS x KISUMU hybrids.

Financial support: This study was supported by a Hellenic Secretariat General for Research and Technology grant awarded to GS (Operational Program Competitiveness, Action 4.3.6.1) and the Innovative Vector Control Consortium (DN and JV).

* Address correspondence to George Skavdis, Department of Molecular Biology and Genetics, Democritus University of Thrace, University Campus, Building 10, Dragana, Alexandroupolis 68100, Greece. E-mail: gskavdis{at}mbg.duth.gr

Authors’ addresses: Athina Kazanidou, Maria Grigoriou, and George Skavdis, Department of Molecular Biology and Genetics, Democritus University of Thrace, University Campus, Building 10, Dragana, Alexandroupolis 68100, Greece, Tel: 0030-2551030610, Fax: 0030-2551030613, E-mails: athina0603{at}yahoo.com, mgrigor{at}mbg.duth.gr, and gskavdis{at}mbg.duth.gr. Dimitra Nikou, Vector Group, Liverpool School of Tropical Medicine, Pembroke Place, Liverpool L35QA, UK, Tel: 0044-1517053156, E-mail: dim_nikou{at}yahoo.co.uk. John Vontas, Laboratory of Pesticide Science, Agricultural University of Athens, Iera Odos 75, 11855 Votanikos, Athens, Greece, Tel: 0030-6932977437, E-mail: vontas{at}aua.gr. Present address of John Vontas, Faculty of Biotechnology and Applied Biology, Department of Biology, University of Crete, Crete, Greece.

Reprint requests: George Skavdis, Democritus University of Thrace, Department of Molecular Biology and Genetics, University Campus, Building 10, Dragana, Alexandroupolis 68100, Greece, E-mail: gskavdis{at}mbg.duth.gr.






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Copyright © 2009 by the American Society of Tropical Medicine and Hygiene.