A mutation in the voltage-gated sodium channel gene associated with pyrethroid resistance in Latin American Aedes aegypti
A mutation in the voltage-gated sodium channel gene associated with pyrethroid resistance in Latin American Aedes aegypti
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DOI：
10.1111/j.1365-2583.2007.00774.x
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发表时间：
2007-12-01
影响因子：
2.6
通讯作者：
Black, W. C.
中科院分区：
文献类型：
--
作者：
Saavedra-Rodriguez, K.;Urdaneta-Marquez, L.;Black, W. C.
拟除虫菊酯通常用作杀成蚊剂，对这些化合物的抗药性的进化是对公共健康的主要威胁。对拟除虫菊酯（kdr）的“击倒抗性”通常是由电压门控钠通道跨膜蛋白（para）中的非同义突变引起的，该突变减少了拟除虫菊酯的结合。早期检测 kdr 对于制定蚊子耐药性管理策略至关重要，其中包括埃及伊蚊（登革热和黄热病病毒最流行的载体）。布伦格斯等人。描述了 Ae 中 para 结构域 II 疏水片段 6 的 7 个新突变。埃及伊蚊。对带有这些突变的菌株的幼虫进行的分析表明，神经对氯菊酯抑制的敏感性降低。其中两个分别发生在外显子 20 和 21 的密码子 Iso1011 和 Val1016 中。 Iso1011 第三个位置的转变编码 Met1011 替换，Val1016 第二个位置的颠换编码 Gly1016 替换。我们在同一区域的 1318 只蚊子中筛查了另外 32 个品系； 30 名来自整个拉丁美洲。虽然在拉丁美洲从未检测到 Gly1016 等位基因，但我们在这些相同的密码子中发现了两个新的突变。密码子 1011 第一个位置的转变编码 Val 替换，而密码子 1016 第一个位置的转变编码 Iso 替换。我们开发了针对这四种突变的 PCR 检测方法，可以在琼脂糖凝胶上读取或作为熔解曲线读取。选择实验中，一项对来自古巴圣地亚哥的野外菌株使用溴氰菊酯，另一项对来自墨西哥女人岛的菌株使用氯菊酯，结果迅速增加了 Iso1016 等位基因的频率。对氯菊酯敏感的 Val1016 纯合父母和氯菊酯抗性 Iso1016 纯合父母产生的 F-3 后代的生物测定表明，Iso1016 在赋予 kdr 时作为隐性等位基因分离。对 F-3 中 1011 和 1016 密码子等位基因之间的分离分析显示，尽管这两个密码子仅被类似的 250 bp 内含子分开，但重组率很高。所提供的工具和信息提供了一种早期检测和表征 kdr 的方法，这对于制定抗性管理策略至关重要。
Pyrethroids are commonly used as mosquito adulticides and evolution of resistance to these compounds is a major threat to public health. 'Knockdown resistance' to pyrethroids (kdr) is frequently caused by nonsynonymous mutations in the voltage-gated sodium channel transmembrane protein (para) that reduce pyrethroid binding. Early detection of kdr is critical to the development of resistance management strategies in mosquitoes including Aedes aegypti, the most prevalent vector of dengue and yellow fever viruses. Brengues et al. described seven novel mutations in hydrophobic segment 6 of domain II of para in Ae. aegypti. Assays on larvae from strains bearing these mutations indicated reduced nerve sensitivity to permethrin inhibition. Two of these occurred in codons Iso1011 and Val1016 in exons 20 and 21 respectively. A transition in the third position of Iso1011 encoded a Met1011 replacement and a transversion in the second position of Val1016 encoded a Gly1016 replacement. We have screened this same region in 1318 mosquitoes in 32 additional strains; 30 from throughout Latin America. While the Gly1016 allele was never detected in Latin America, we found two new mutations in these same codons. A transition in the first position of codon 1011 encodes a Val replacement while a transition in the first position of codon 1016 encodes an Iso replacement. We developed PCR assays for these four mutations that can be read either on an agarose gel or as a melting curve. Selection experiments, one with deltamethrin on a field strain from Santiago de Cuba and another with permethrin on a strain from Isla Mujeres, Mexico rapidly increased the frequency of the Iso1016 allele. Bioassays of F-3 offspring arising from permethrin susceptible Val1016 homozygous parents and permethrin resistant Iso1016 homozygous parents show that Iso1016 segregates as a recessive allele in conferring kdr. Analysis of segregation between alleles at the 1011 and 1016 codons in the F-3 showed a high rate of recombination even though the two codons are only separated by a similar to 250 bp intron. The tools and information presented provide a means for early detection and characterization of kdr that is critical to the development of strategies for resistance management.