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A New Robust Diagnostic Polymerase Chain Reaction for Determining the Mating Status of Female Anopheles gambiae Mosquitoes

The principal malaria vector in Africa, Anopheles gambiae, contains two pairs of autosomes and one pair of sex chromosomes. The Y chromosome is only associated with males and other Y chromosome–specific DNA sequences, which are transferred to women during mating. A reliable tool to determine the mating status of dried wild An. gambiae females is currently lacking. DNA was extracted from dried virgin and mated females and used to test whether Y chromosome–specific polymerase chain reaction (PCR) markers can be successfully amplified and used as a predictor of mating. Here we report a new PCR-based method to determine the mating status among successfully inseminated and virgin wild An. gambiae females, using three male-specific primers. This dissection-free method has the potential to facilitate studies of both population demographics and gene flow from dried mosquito samples routinely collected in epidemiologic monitoring and aid existing and new malaria-vector control approaches.

Received January 31, 2007. Accepted for publication April 30, 2007.

Acknowledgments: We thank the Vector Genetic Laboratory (University of California) for cooperation during the entire period of this study; Claudio Meneses for invaluable technical assistance during mosquito rearing and PCR optimization; Dr. Mark Benedict for suggestions and literature that made this study possible; and Dr. Heather Ferguson for reviewing the manuscript before submission.

Financial support: We acknowledge the International Atomic Energy Agency (IAEA) for financial support through a fellowship awarded to KRN to work at UC Davis. This research forms part of a VIDI Grant 864.03.004 from the Dutch Scientific Organization (NWO) awarded to BGJK. In addition, we acknowledge support to GCL through Grant AI040308 from the National Institutes of Health.

^* Address correspondence to Kija R. Ng’habi, Ifakara Health Research and Development Centre, PO Box 53, Ifakara, United Republic of Tanzania. E-mail: kija{at}ihrdc.or.tz

Authors’ addresses: Kija R Ng’habi, Ifakara Health Research and Development Centre, PO Box 53, Ifakara, United Republic of Tanzania, E-mail: kija{at}ihrdc.or.tz. Ashley Horton and Gregory Lanzaro, Department of Entomology, University of California, Davis, 396C Briggs, One Shield Avenue, Davis, CA 95616-8584, E-mails: gclanzaro{at}ucdavis.edu and ahorton{at}ucdavis.edu. Bart G. J. Knols, Laboratory of Entomology. PO Box 8031, 6700 EH, Wageningen University and Research Centre, Wageningen, The Netherlands, E-mail: Bart.Knols{at}wur.nl.