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生物素DNA探针/PCR系统用于班氏丝虫蚊媒监测的研究

Detection of Filarial Larvae in Mosquitoes with Wuchereria Bancrofti Specific DNA Probe Labeled with Biotin/PCR System

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【作者】 黄炳成常江韩广东张玉林刘凤梅王士华陈锡欣王凯甄天民李连云贾凤菊刘玉俊程义亮

【Author】 Huang Bingcheng * Chang Jiang Han Guangdong et al (* Shandong Institute of Parasitic Diseases,Jining 272133)

【机构】 山东省寄生虫病防治研究所!济宁272033河南省卫生防疫站河南省?

【摘要】 目的 :评价班氏丝虫特异生物素DNA探针 /PCR系统用于丝虫病防治后期蚊媒监测的效果。方法 :应用长臂光敏生物素标记的班氏丝虫种特异的DNA片段 (4 90bp)为探针 ,结合聚合酶链反应技术 ,对实验室感染蚊虫和现场捕获蚊虫进行检测。结果 :该探针只与班氏丝虫感染蚊提取的DNA杂交 ,不与马来丝虫感染蚊提取的DNA、其他动物丝虫DNA以及正常蚊DNA杂交。同批人工感染班氏微丝蚴蚊虫 ,人工解剖镜检阳性率为 2 1.2 8% ,探针检测阳性率为 30 .6 7% ,差异无显著性 (P >0 .0 5)。在平均感染度为 1.6条幼虫 /蚊情况下 ,50只蚊虫中有 1只感染蚊即可被探针检出。在河南省班氏丝虫病流行区自病家和病村其他户分别捕获蚊虫 ,应用探针每 2 0只为一组进行检测 ,蚊虫最低阳性率分别为 0 .75%和 0 .19% ,检测非病村正常蚊均为阴性。结论 :该DNA探针 /PCR系统的敏感性和特异性均达到实用水平 ,为丝虫病防治后期的监测增添了一新的有用工具

【Abstract】 AIM: To evaluate the usefulness of biotin-DNA probe/PCR system for post control surveillance of Bancroftian filariasis in mosquito. Methods: This assay was used to detect the mosquitoes infected in a laboratory and in an endemic area by the way of W.bancrofti specific DNA probe labelled with photobiotin, after samples amplified by PCR. Results: This probe hybridized with DNA of W.bancrofti only, not with DNAs of B. malayi , some animal filariae and normal mosquitoes. Among the same batch mosquitoes artificially infected with W. bancrofti , the positive rates were 30.67 % by probe, and 21.28 % by dissectting microscopy. Under infectiosity of 1.6 filarial larvae/mosquito, it was detectable when 1 out of 50 mosquitoes was infected. When each group with 20 mosquitoes was detected by the system of DNA probe/PCR system, the lowest positive rates of mosquitoes caught from patients and non patientshouse in the same village of Henan province( W.bancrofti endemic area) were 0.75 % and 0.19 % respectively, but the mosquitoes caught from non patient village were all negative. Conclusion: Results show that the method had reached the practical level because its high sensitivity and specifty and also proved a new useful tool for the surveillance of filariasis in the late stage control.

  • 【文献出处】 中国媒介生物学及控制杂志 ,CHINESE JOURNAL OF VECTOR BIOLOGY AND CONTROL , 编辑部邮箱 ,2000年03期
  • 【分类号】R181.8
  • 【被引频次】4
  • 【下载频次】77
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