Frequency Detection of Organophosphate Resistance Allele in Anopheles sinensis (Diptera: Culicidae) Populations by Real-time PCR Amplification of Specific Allele (rtPASA) |
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| Affiliation: | 1. Biology Center of the Czech Academy of Sciences, Branisovska 31, 37005 Budweis, (Ceske Budejovice), Czechia;2. Dpto. de Genética, Facultad de Ciencias, Universidad de Granada, Campus de Fuentenueva s/n, 18071, Granada, Spain;3. Lab. de Bioinformática, Centro de Investigación Biomédica (CIBM), PTS, Avda. del Conocimiento s/n, 18100, Granada, Spain;1. Odawara Research Center, Nippon-soda Co., Ltd., 345 Takada, Odawara, Kanagawa 250-0216, Japan;2. Insect Genome Research and Engineering Unit, Division of Applied Genetics, Institute of Agrobiological Sciences, NARO, 1-2 Owashi, Tsukuba, Ibaraki 305-8634, Japan;1. Research Institute of Agriculture and Life Sciences, Seoul National University, Seoul 08826, Republic of Korea;2. EntoCode Co., Seoul 06028, Republic of Korea;3. Biological and Genetic Resources Assessment Division, National Institute of Biological Resources, Incheon 22689, Republic of Korea;4. Department of Agricultural Biology, Seoul National University, Seoul 08826, Republic of Korea;1. Department of Biology, Faculty of Science, Mugla Sitki Kocman University, Mugla, Turkey;2. Koycegiz Vocational School, Department of Medicinal and Aromatic Plants, Mugla Sitki Kocman University, Mugla, Turkey;1. Department of Agricultural Biology, Seoul National University, Sillim 9-dong, Seoul, Republic of Korea;2. Ilsong Institute of Life Science, Hallym University, Anyang, Gyeonggido, Republic of Korea;3. Department of Biomedical Gerentology, Hallym University, Chuncheon, Gangwondo, Republic of Korea;4. Korea Basic Science Institute, Sungbuk-gu, Seoul, Republic of Korea;5. Department of Obstetrics and Gynecology, Hallym University Sacred Heart Hospital, Anyang, Gyeonggido, Republic of Korea;6. National Institute of Biological Resources, Environmental Research Complex, Gyoungseo-dong, Seo-gu, Incheon, Republic of Korea;7. Research Institute for Agriculture and Life Science, Seoul National University, Sillim 9-dong, Seoul, Republic of Korea |
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| Abstract: | A rapid, simple and accurate real-time PASA (PCR amplification of specific allele) (rtPASA) protocol was developed and optimized for the frequency estimation of the Glyl 19Ser mutation in the type-I acetylcholinesterase locus, putatively associated with organophosphate resistance, in pooled DNA samples of Anopheles sinensis, a major vector mosquito of malaria in Korea. Performance of the rtPASA protocol was evaluated by comparing with the data generated from individual genotypings of a field population. The resistance allele frequency of the population (74.4%) predicted from the linear regression line of the rtPASA agreed well with that estimated from the individual genotyping (74.1%), demonstrating its reliability and accuracy. Using this rtPASA protocol, the resistance allele frequency in 10 local populations of An. sinensis was determined to range from 74.4% to 97.2%, suggestive of the widespread organophosphate resistance in An. sinensis in Korea. |
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