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A novel strategy for screening mutations in the voltage-gated sodium channel gene of Aedes albopictus based on multiplex PCR-mass spectrometry minisequencing technology

查看全文 作　　者：Qunzheng [1,2]Mu;Xin [1]Zhao;Fengfeng [3]Li;Wenyu [4]Li;Xinxin [1]Zhou;Xinchang [1]Lun;Yiguan [1]Wang;Dongdong [1]Hua;Qiyong [1]Liu;Di [1]Xiao;Fengxia [1]Meng 高影响力作者 机构地区：[1]National Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases,National Institute for Communicable Disease Control and Prevention,Chinese Center for Disease Control and Prevention,Beijing,102206,People’s Republic of China;[2]Weifang No.2 People’s Hospital,Weifang,261000,Shandong,People’s Republic of China;[3]Weifang Medical College,Weifang,261000,Shandong,People’s Republic of China;[4]Beijing Daxing District Center for Disease Control and Prevention,Beijing,102600,Beijing,People’s Republic of China高影响力机构 出　　处：《Infectious Diseases of Poverty》索引2023年第12卷第4期,共11页高影响力期刊 基　　金：National Science and Technology Major Project of China(No. 2018ZX10101002-002)。 摘　　要：Background The current prevention and control strategy forAedes albopictus heavily relies on comprehensive management, such as environmental management and chemical control. However, the wide application of pyrethroids has facilitated the development of insecticide resistance, primarily via mutations in the voltage-gated sodium channel (VGSC) gene. This study aims to develop a novel strategy for detecting mutations in the VGSC gene inAe. albopictus using multiplex PCR-mass spectrometry (MPCR-MS) minisequencing technology.Methods We established a new strategy for detecting mutations in the VGSC gene inAe. albopictus using MPCR-MS minisequencing technology. MPCR amplification and mass probe extension (MPE) were first used, followed by single nucleotide polymorphism (SNP) typing mass spectrometry, which allows the simultaneous detection of multiple mutation sites of the VGSC gene in 96 samples ofAe. albopictus. A total of 70 wild-collectedAe. albopictus were used to evaluate the performance of the method by comparing it with other methods.Results Three target sites (1016, 1532, 1534) in the VGSC gene can be detected simultaneously by double PCR amplification combined with matrix-assisted laser desorption ionization-time-of-flight mass spectrometry, achieving a detection limit of 20 fg/μl. We applied this method to 70 wild-collectedAe. albopictus, and the obtained genotypes were consistent with the routine sequencing results, suggesting the accuracy of our method.Conclusions MPCR-MS minisequencing technology provides a sensitive and high-throughput approach toAe. albopictus VGSC gene mutation screening. Compared with conventional sequencing, this method is economical and time-saving. It is of great value for insecticide resistance surveillance in areas with a high risk of vector-borne disease. 关 键 词：Aedes albopictus VGSC gene MUTATION Single nucleotide polymorphisms Multiplex polymerase chain reaction-mass spectrometry mini-sequencing
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