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目的了解山东省不同地区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。方法采用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。以kdr基因L1014F和L1014S等位基因频率x与成蚊接触筒的存活率y建立的回归方程分别为y=1.130566x+0.34394(r2=0.811684,P<0.05)和y=2.919579x+1.06308(r2=0.997039,P<0.05)。结论淡色库蚊溴氰菊酯抗药性与kdr基因突变呈正相关,具体的抗药性机理亟待进一步研究。
Objective To understand the resistance level of deltamethrin to adult mosquito Culex pipiens pallens in different areas of Shandong Province and the relationship between the mutation and drug resistance of kdr allele. Methods The kdr gene sequence of adult Culex pipiens pallens from different regions was cloned by PCR 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); L104S: TTA is mutated to TCA and the corresponding leucine (L) is replaced by serine (S). The regression equations established with the frequencies of allele k of kdr gene L1014F and L1014S x and the survival rate of adult mosquitoes were y = 1.130566x + 0.34394 (r2 = 0.811684, P <0.05) and y = 2.919579x + 1.06308 = 0.997039, P <0.05). Conclusion The resistance to deltamethrin in Culex pipiens pallens is positively correlated with the mutation of kdr gene. The specific drug resistance mechanism needs to be further studied.