title: First detection of multiple knockdown resistance (kdr)-like mutations in voltage-gated sodium channel using three new genotyping methods in Anopheles sinensis from Guangxi Province, China.
abstract: To investigate knockdown resistance (kdr)-like mutations associated with pyrethroid resistance in Anopheles sinensis (Wiedemann, 1828), from Guangxi province, southwest China, a segment of a sodium channel gene was sequenced and genotyped using three new genotyping assays. Direct sequencing revealed the presence of TTG-to-TCG and TG-to-TTT mutations at allele position L1014, which led to L1014S and L1014F substitutions in a few individual and two novel substitutions of N1013S and L1014W in two DNA templates. A low frequency of the kdr allele mostly in the heterozygous state of L1014S and L1014F was observed in this mosquito population. In this study, the genotyping of An. sinensis using three polymerase chain reaction-based methods generated consistent results, which agreed with the results of DNA sequencing. In total, 52 mosquitoes were genotyped using a direct sequencing assay. The number of mosquitoes and their genotypes were as follows: L/L = 24, L/S = 19, L/F = 8, and F/W = 1. The allelic frequency of L1014, 1014S, and 1014F were 72, 18, and 9%, respectively.
authors.affiliation: State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, 20 Fengtai East St, Beijing 100071, PR China.
authors.forename: Wei L
authors.initials: WL
authors.lastname: Tan
authors.forename: Chun X
authors.initials: CX
authors.lastname: Li
authors.forename: Zhong M
authors.initials: ZM
authors.lastname: Wang
authors.forename: Mei D
authors.initials: MD
authors.lastname: Liu
authors.forename: Yan D
authors.initials: YD
authors.lastname: Dong
authors.forename: Xiang Y
authors.initials: XY
authors.lastname: Feng
authors.forename: Zhi M
authors.initials: ZM
authors.lastname: Wu
authors.forename: Xiao X
authors.initials: XX
authors.lastname: Guo
authors.forename: Dan
authors.initials: D
authors.lastname: Xing
authors.forename: Ying M
authors.initials: YM
authors.lastname: Zhang
authors.forename: Zhong C
authors.initials: ZC
authors.lastname: Wang
authors.forename: Tong Y
authors.initials: TY
authors.lastname: Zhao
mesh_terms: D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000818:Animals; D000852:Anopheles / Q000235:genetics*; D001483:Base Sequence; D002681:China; D005260:Female; D060005:Genotyping Techniques; D007305:Insecticide Resistance / Q000235:genetics*; D008969:Molecular Sequence Data; D009154:Mutation; D016133:Polymerase Chain Reaction; D015222:Sodium Channels / Q000235:genetics*
chemical_list: D015222:Sodium Channels