论文部分内容阅读
目的了解山东省不同地区的淡色库蚊成蚊对溴氰菊酯的抗药性水平及其kdr等位基因的突变与抗药性水平间的关联情况,并探索建立蚊虫抗药性的现场检测方法。方法采用PCR和AS-PCR技术,使用特异性引物克隆不同地区野生淡色库蚊成蚊kdr基因序列,并进行基因测序。利用线性回归分析判断其突变情况与抗药性的关联情况。结果成功克隆出kdr等位基因,通过基因序列分析发现在淡色库蚊钠通道第II结构域S6节段1014位点上存在2种突变,即L1014F:TTA突变为TTT,相应的亮氨酸(L)被苯丙氨酸(F)取代,L1014S:TTA突变为TCA,相应的亮氨酸(L)被丝氨酸(S)取代。Kdr基因L1014F突变和L1014S突变与淡色库蚊对溴氰菊酯的抗性表型呈正相关。蚊虫抗药性检测的结果显示PCR技术优于AS-PCR技术。结论淡色库蚊kdr基因突变与溴氰菊酯抗药性表型呈正相关,PCR和AS-PCR技术都可以应用于蚊虫抗药性的现场检测。
Objective To understand the relationship between resistance to deltamethrin and the mutation and drug resistance of kdr alleles in adults of Culex pipiens pallens in different areas of Shandong Province and explore the establishment of on-site detection methods for mosquito resistance. Methods The kdr gene sequences of adult Culex pipiens pallens from different regions were cloned by PCR and AS-PCR techniques and sequenced. Using linear regression analysis to determine the relationship between the mutation and drug resistance. Results The kdr allele was cloned successfully. Two mutations were found in the S6 segment of the second domain of Culex pipiens pallens, ie L1014F: TTA was mutated to TTT and the corresponding leucine ( L) is replaced by phenylalanine (F), L1014S: TTA is mutated to TCA, and the corresponding leucine (L) is replaced by serine (S). The L1014F mutation and L1014S mutation of Kdr gene were positively correlated with resistance phenotypes of deltamethrin in Culex. Mosquito resistance test results show that PCR technology is superior to AS-PCR technology. Conclusion The mutation of kdr gene in Culex pipiens pallens is positively correlated with the resistance phenotype of deltamethrin. Both PCR and AS-PCR techniques can be applied to the field detection of mosquito resistance.