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MULTIPLEX PCR ASSAY FOR THE DETECTION OF ANOPHELES FLUVIATILIS SPECIES COMPLEX, HUMAN HOST PREFERENCE, AND PLASMODIUM FALCIPARUM SPOROZOITE PRESENCE, USING A UNIQUE MOSQUITO PROCESSING METHOD

A multiplex PCR assay has been developed for detection of Anopheles fluviatilis cryptic species, their human host preference, and Plasmodium falciparum presence in the mosquito. PCR conditions were optimized using primer sets specific for A. fluviatilis cryptic species, Homo sapiens, and P. falciparum and evaluated with field-collected mosquitoes. A unique mosquito processing method was used for screening P. falciparum carrying capacity and human host preference of A. fluviatilis mosquitoes in first-round multiplex PCR. The vectorial status of the mosquito for P. falciparum parasite was confirmed in second-round PCR. Of the 121 collected mosquitoes, 92 were of S type, 26 of T type, and 3 were of other types. Human host preference was dominant in S type, of which 4% were P. falciparum sporozoite positive. This assay and processing method can also be used to evaluate vector competence of other anophelines.

Received August 7, 2006. Accepted for publication December 13, 2006.

Acknowledgments: The authors thank Dr. B. Ravindran, Director of the Institute of Life Sciences, Bhubaneswar, India, for his support. We are grateful to Mr. H.K. Tripathy and Mr. N.S. Marai for technical help and to the persons who maintain the insectary and animal facility at Regional Medical Research Centre, Bhubaneswar, India.

Financial support: This work was supported by funds from Department Biotechnology, Government of India. Mr. Amitav Mohanty was supported by the fellowship from the Council of Scientific and Industrial Research, Government of India.

^* Address correspondence to Rupenangshu K. Hazra, Regional Medical Research Centre, Nalco Square, Chandrasekharpur, Bhubaneswar 751 023, Orissa, India. E-mail: rupenkh{at}yahoo.co.in

Authors’ addresses: Amitav Mohanty, Priyanka Kar, Kirti Mishra, and Durg V. Singh, Division of Vector Borne Diseases, Institute of Life Sciences, Nalco Square Chandrasekharpur, Bhubaneswar 751 023, Orissa, India, Telephone: +91-6742301900, Fax: +91-6742300728, E-mail: amitav{at}ils.res.in. Namita Mohapatra, Santanu K. Kar, and Rupenangshu K. Hazra, Regional Medical Research Centre, Nalco Square, Bhubaneswar 751 023, Orissa, India, Telephone: +91-6742301416, +91-9861173867, Fax: +91-6742301351, E-mail: rupenkh{at}yahoo.co.in. Aditya P. Dash, National Institute of Malaria Research, 22 Shamnath Marg, Delhi 110054, India, Telephone: +91-1122533201, Fax: +91-1122515086.

Reprint requests: Rupenangshu K. Hazra, Regional Medical Research Centre, Nalco Square, Bhubaneswar 751 023, Orissa, India. Telephone: +91-6742301416, +91-9861173867, Fax: +91-6742301351, E-mail: rupenkh{at}yahoo.co.in.