Molecular ecology of pyrethroid knockdown resistance in Culex pipiens pallens mosquitoes.
Molecular ecology of pyrethroid knockdown resistance in Culex pipiens pallens mosquitoes.
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DOI：
10.1371/journal.pone.0011681
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发表时间：
2010-07-21
期刊：
影响因子：
3.7
通讯作者：
Yan G
中科院分区：
文献类型：
--
作者：
Chen L;Zhong D;Zhang D;Shi L;Zhou G;Gong M;Zhou H;Sun Y;Ma L;He J;Hong S;Zhou D;Xiong C;Chen C;Zou P;Zhu C;Yan G
拟除虫菊酯类杀虫剂在中国和世界范围内广泛用于公共卫生害虫控制。准确的抗药性监测对于指导杀虫剂的合理使用和抗药性管理至关重要。在这里，我们研究了对位钠通道基因的核苷酸多样性，这赋予敲低抗性（kdr）在库蚊在中国的pipiens pallens。对钠离子通道基因的序列分析确定了L1014 F和L1014 S突变。我们开发并验证了等位基因特异性PCR和实时TaqMan方法用于抗性诊断。实时荧光定量TaqMan方法的扩增率、灵敏度和特异性均优于等位基因特异性PCR方法，具有上级优势。kdr等位基因频率与生物测定抗性表型之间存在显著正相关，表明kdr基因L1014F和L1014S突变频率可作为天然Cx溴氰菊酯抗性监测的分子标记。pipiens pallens在华东地区的分布。室内选择试验发现，L1014 F突变频率对溴氰菊酯选择有响应，而L1014 S突变频率对溴氰菊酯选择无响应，表明L1014 F突变是产生溴氰菊酯抗性的关键突变。在六个Cx群体中检测到高L1014F突变频率。pipens pallens表明中国东部的拟除虫菊酯耐药性很高，需要进一步调查，以绘制中国的耐药性地图，并研究替代的蚊虫控制策略。
Pyrethroid insecticides have been extensively used in China and worldwide for public health pest control. Accurate resistance monitoring is essential to guide the rational use of insecticides and resistance management. Here we examined the nucleotide diversity of the para-sodium channel gene, which confers knockdown resistance (kdr) in Culex pipiens pallens mosquitoes in China. The sequence analysis of the para-sodium channel gene identified L1014F and L1014S mutations. We developed and validated allele-specific PCR and the real-time TaqMan methods for resistance diagnosis. The real-time TaqMan method is more superior to the allele-specific PCR method as evidenced by higher amplification rate and better sensitivity and specificity. Significant positive correlation between kdr allele frequency and bioassay-based resistance phenotype demonstrates that the frequency of L1014F and L1014S mutations in the kdr gene can be used as a molecular marker for deltamethrin resistance monitoring in natural Cx. pipiens pallens populations in the East China region. The laboratory selection experiment found that L1014F mutation frequency, but not L1014S mutation, responded to deltamethrin selection, suggesting that the L1014F mutation is the key mutation conferring resistance to deltamethrin. High L1014F mutation frequency detected in six populations of Cx. pipens pallens suggests high prevalence of pyrethroid resistance in Eastern China, calling for further surveys to map the resistance in China and for investigating alternative mosquito control strategies.
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