title: Identification of resistant carboxylesterase alleles in Culex pipiens complex via PCR-RFLP.
abstract: BACKGROUND
  Carboxylesterase overproduction is a frequently observed resistance mechanism of insects to organophosphate insecticides. As a major transmitter of human diseases, mosquitoes in the Culex pipiens complex have evolved 13 carboxylesterase alleles (Ester) that confer organophosphate resistance. Six alleles, Ester(B1), Ester², Ester⁸, Ester⁹, Ester(B10), and Ester¹¹, have been observed in field populations in China, sometimes co-existing in one population. To differentiate the carboxylesterase alleles found in these field populations, PCR-RFLP was designed for use in resistance monitoring.
  
  
  RESULTS
  Based on the DNA sequences of resistant and nonresistant carboxylesterase alleles, Ester B alleles were first amplified with PCR-specific primers and then digested with the restriction enzyme DraI. In this step, Ester² and Ester¹¹ were differentiated from the other Ester alleles. When the other Ester B alleles were digested with the restriction enzyme XbaI, Ester(B1) and the susceptible C. p. pallens Ester were screened out. Ester⁸ and Ester⁹ were differentiated from Ester(B10) and the susceptible C. p. quinquefasciatus esterase allele, respectively, by amplifying and digesting the Ester A alleles with the restriction enzyme ApaLI. The effectiveness of the custom-designed PCR-RFLP was verified in two field mosquito populations.
  
  
  CONCLUSIONS
  A PCR-RFLP based approach was developed to differentiate carboxylesterase alleles in Culex pipiens complex mosquitoes. These processes may be useful in monitoring the evolutionary dynamics of known carboxylesterase alleles as well as in the identification of new alleles in field populations.
authors.affiliation: State Key Laboratory of Integrated Management of Pest Insects and Rodents, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China.
authors.forename: Hanying
authors.initials: H
authors.lastname: Zhang
authors.forename: Fengxia
authors.initials: F
authors.lastname: Meng
authors.forename: Chuanling
authors.initials: C
authors.lastname: Qiao
authors.forename: Feng
authors.initials: F
authors.lastname: Cui
mesh_terms: D000483:Alleles; D000818:Animals; D043182:Carboxylesterase / Q000235:genetics*; D002681:China; D003465:Culex / Q000187:drug effects* / Q000201:enzymology*; D004199:Disease Vectors; D004772:Entomology / Q000379:methods; D007305:Insecticide Resistance; D010755:Organophosphates / Q000378:metabolism / Q000494:pharmacology*; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length
chemical_list: D010755:Organophosphates; D043182:Carboxylesterase