title: [Study on TaqMan-MGB real-time fluorescence quantitative PCR to detect gene mutation of kdr from Anopheles sinensis].
abstract: OBJECTIVE
  To establish a Real-time Fluorescence Quantitative PCR to detect the kdr gene mutation in Anopheles sinensis.
  
  
  METHODS
  One pair of primers and three TaqMan-MGB probes were designed based on kdr gene and its L1014 locus mutations of A. sinensis. After optimization, the Real-time Fluorescence Quantitative PCR was verified by using 6 types of A. sinensis samples with different kdr gene types. Additionally, 50 laboratory samples and 113 field samples were tested by this method.
  
  
  RESULTS
  The established Real-time Fluorescence Quantitative PCR could identify 6 different kdr gene types in A. sinensis. The mutation could be detected by single-tube Fluorescence Quantitative PCR, and the detail mutation type could be further identified by double-tube Fluorescence Quantitative PCR. By using this method, 50 laboratory samples were confirmed as wild type homozygotes. Among 113 field samples, 12 were wild type homozygotes, others were L1014F or L1014C mutations, and the total mutation frequency was 87.61%.
  
  
  CONCLUSIONS
  The new established TaqMan-MGB Real-time Fluorescence Quantitative PCR can be used to detect the kdr gene L1014 mutations of A. sinensis.
authors.affiliation: Jiangsu Institute of Parasitic, Key Laboratory on Technology for Parasitic Disease Prevention and Control, Ministry of Health, Wuxi 214064, China.
authors.forename: Liang
authors.initials: L
authors.lastname: Bai
authors.forename: Guo-Ding
authors.initials: GD
authors.lastname: Zhu
authors.forename: Jian-Xia
authors.initials: JX
authors.lastname: Tang
authors.forename: Chao
authors.initials: C
authors.lastname: Zhang
authors.forename: Yao-Bao
authors.initials: YB
authors.lastname: Liu
authors.forename: Ju-Lin
authors.initials: JL
authors.lastname: Li
authors.forename: Jun
authors.initials: J
authors.lastname: Cao
authors.forename: Qi
authors.initials: Q
authors.lastname: Gao
mesh_terms: D000818:Animals; D000852:Anopheles / Q000235:genetics*; D001483:Base Sequence; D017931:DNA Primers; D008969:Molecular Sequence Data; D009154:Mutation; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*
chemical_list: D017931:DNA Primers