敌敌畏_____淡色库蚊；敌敌畏；(临床诊治)

敌敌畏抗性淡色库蚊酯酶B1、A2探针制备及应用

http://www.100md.com 《中国热带医学杂志》 2005年第2期

敌敌畏,,淡色库蚊；敌敌畏；,酯酶B1、A2；DNA探针

     摘要：目的制备敌敌畏抗性品系淡色库蚊酯酶B1、A2探针，并应用于抗性品系及敏感品系的检测。方法分别设计酯酶B1、A2上下游引物，5’端地高辛标记，PCR法制备DNA探针，分子杂交检测各个品系淡色库蚊。结果摸索出合适的区分抗性与敏感品系的探针浓度，在该浓度下各品系杂交点的阳性率有差别。应用酯酶B1探针检测各品系蚊虫，抗敌敌畏品系的阳性率为40.0%，抗残杀威品系的阳性率为23.3%，抗氯氰菊酯品系的阳性率为183%, 敏感品系的阳性率为117%；应用酯酶A2探针检测各品系蚊虫，抗敌敌畏品系的阳性率为433%，抗残杀威品系的阳性率为250%，抗氯氰菊酯品系的阳性率为217%, 敏感品系的阳性率为150%。结论应用酯酶B1、A2探针能够区分抗性品系，因此能够用于抗性的测定。从而为抗有机磷类杀虫剂媒介昆虫基因检测的实用性和抗性分子生物学机制的研究打下基础。

    关键词：淡色库蚊；敌敌畏；酯酶B1、A2；DNA探针

    中图分类号：R3841 文献识别码：A 文章编号：1009-9727(2005)02-193-03

    Preparation and application of esterase B1 and A2 probe for detection of the resistance of antiDDVP Culex pipiens pallens CUI Wei1, ZHEN Tian-min1, WANG Huai-wei1, et al (1Shandong Provincial Institute of Parasitic Diseases, Jining 272033, Shandong, P R China； 2Inistitute of Basic Medicine,Shandong Academy of Medical SciencesJinan 250062,Shandong,PRChina)

    Abstract：Objective To prepare the esterase B1、A2 probe of DDVP resistant Culex pipiens pallens and detect resistant level of Culex pipiens pallens Methods ForwardPrimer and ReversePrimer of esterase B1、A2, were designed and labeled with digoxin on 5’ end of ForwardPrimer of esterase B1、A2 DNA probe was prepared by PCR and used for identification and screening the DDVP resistance of Culex pipiens pallens Results The probe concentration suitable for the differentiation of resistant and sensitive strains were ascertained by repeated experiment In this concentration, the positive rate of hybrid were different among all strains It shows that with use of esterase Bl probe the positive rates of the strains resistant to DDVP,proproxur and permethrin were 40%,23% and 183%,respectively,while by use of esterase A2 probe the positive rates of the strains to DDVP,propoxur and permethrin were 43%,25% and 117%,respectively Conclusion The different resistant strains can be distinguished with esterase B1 probe and esterase A2 probe, thereby we can apply it to detect every sesistant strains This provided a practical molecular biological method for gene detection of DDVP resistant vector insects ......

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