Rasli Rosilawati, Lee Han Lim, Wasi Ahmad Nazni, Fikri Siti Futri Farahininajua, Ali Roziah, Muhamed Khairul Asuad, Hadi Azahari Abdul, Liu Qi-Yong, Meng Feng Xia
Medical Entomology Unit, Infectious Disease Research Centre, Institute for Medical Research, Jalan Pahang, Kuala Lumpur 505888, Malaysia.
State Key Laboratory for Infectious Diseases Prevention and Control, National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China.
Insects. 2018 Apr 18;9(2):43. doi: 10.3390/insects9020043.
This study is intended to provide a comprehensive characterization of the resistance mechanisms in the permethrin-selected (IMR-PSS) and laboratory susceptible (IMR-LS) strain from Malaysia. Both IMR-PSS and IMR-LS provide a standard model for use in assessing the pyrethroid resistance in field-collected strains collected from three dengue hotspots: the Taman Seri Bayu (TSB), the Flat Camar (FC), and the Taman Dahlia (TD). Two established methods for determining the resistance mechanisms of the pyrethroid are the quantification of detoxification enzymes via enzyme microassay and the nucleotide sequencing of the domain 2 region from segment 1 to 6 via classical polymerase chain reaction (PCR) amplification-were employed. Enzyme activities in IMR-LS served as the resistance threshold reference, providing a significant standard for comparison with IMR-PSS and other field-collected strains. The amino acids in the domain 2 region of voltage-gated sodium channel () of IMR-LS were served as the reference for detection of any changes of the knockdown resistance () alleles in IMR-PSS and field-collected strains. Studies clearly indicated that the IMR-LS was highly susceptible to insecticides, whilst the IMR-PSS was highly resistant to pyrethroids and conferred with two resistance mechanisms: the elevated oxidase enzyme activity and the altered target-site mutations. Mutations of V1023G alone, and the combination mutations of V1023G with S996P in IMR-PSS, as well as the in field-collected strain, indicate the spread of the () gene in , particularly in dengue-endemic areas in Malaysia.
本研究旨在全面表征来自马来西亚的氯菊酯选择品系(IMR - PSS)和实验室敏感品系(IMR - LS)中的抗性机制。IMR - PSS和IMR - LS均为标准模型,用于评估从三个登革热热点地区采集的野外品系中的拟除虫菊酯抗性,这三个地区分别是斯里巴尤花园(TSB)、卡马尔公寓(FC)和大丽花花园(TD)。采用了两种既定的测定拟除虫菊酯抗性机制的方法,即通过酶微测定法对解毒酶进行定量,以及通过经典聚合酶链反应(PCR)扩增对第1至6节段的结构域2区域进行核苷酸测序。IMR - LS中的酶活性作为抗性阈值参考,为与IMR - PSS和其他野外采集品系进行比较提供了重要标准。IMR - LS的电压门控钠通道()结构域2区域中的氨基酸用作检测IMR - PSS和野外采集品系中击倒抗性()等位基因任何变化的参考。研究清楚地表明,IMR - LS对杀虫剂高度敏感,而IMR - PSS对拟除虫菊酯高度抗性,并具有两种抗性机制:氧化酶活性升高和靶位点突变改变。IMR - PSS中单独的V1023G突变以及V1023G与S996P的组合突变,以及野外采集的品系中的突变,表明()基因在中的传播,特别是在马来西亚的登革热流行地区。