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Molecular analysis of knock down resistance (kdr) mutation and distribution of kdr genotypes in a wild population of Culex quinquefasciatus from India
Authors:Manas Sarkar  Aparajita Borkotoki  Indra Baruah  Indra Kumar Bhattacharyya   Ravi Bihari Srivastava
Affiliation: Medical Entomology Division, Defence Research Laboratory, Tezpur, India;
 Department of Zoology, Gauhati University, Guwahati, India;
 Cotton College, Guwahati, India;
 Directorate of Life Science, Defence Research &Development Organization, New Delhi, India
Abstract:Objective  To investigate the presence of knock down resistance ( kdr ) mutation, its frequency distribution in the principal vector of bancroftian filariasis, Culex quinquefasciatus from northeastern India, and to relate kdr genotypes with susceptibility and/or resistance to DDT and deltamethrin in this vectors.
Methods  Adult female mosquitoes were collected by aspiration from human dwellings in two villages, Benganajuli and Rikamari, and two military establishments, Field Units I and II. Insecticide susceptibility tests were performed following WHO methods with 4% DDT and 0.05% deltamethrin. Molecular identification of kdr mutation and genotyping of kdr locus was performed by allele-specific PCR (AS-PCR) and direct sequencing in a subset of samples.
Results  Mosquitoes were resistant to DDT and showed 11.9–41.2% mortality, whereas the knock down bioassay for deltamethrin suggests complete susceptibility to this insecticide in all study sites except Benganajuli. The result of AS-PCR confirmed the presence of three genotypes: susceptible (SS), resistant (RR) and heterozygous (SR) in the population. Genotype frequencies at kdr locus for DDT-resistant individuals conformed with the Hardy–Weinberg proportion, whereas DDT and deltamethrin susceptible individuals differed significantly ( P  < 0.05). The efficacy of AS-PCR in detecting the correct genotype was not encouraging.
Conclusions  This is the first report from India on kdr genotyping in C. quinquefasciatus , and it confirms the occurrence of kdr allele in this vector in northeastern India. This finding has serious implications for the filariasis control programmes in India.
Keywords:Culex quinquefasciatus    filariasis    kdr genotype    India    knock down resistance    pyrethroid
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