摘要
为寻找一种敏感、特异的丝虫病监测方法，根据班氏丝虫ｐ Ｗ ｂ１２ 重复序列，用聚合酶链反应扩增班氏丝虫特异的 Ｄ Ｎ Ａ 片段（４９０ ｂｐ），回收后用光敏生物素标记该片段，制备成非放射性 Ｄ Ｎ Ａ 探针，并在探针的制备方法上做了新的尝试。实验结果显示，该探针只与班氏微丝蚴及其感染蚊 Ｄ Ｎ Ａ 出现杂交，与马来微丝蚴、正常蚊以及人白细胞的 Ｄ Ｎ Ａ 未出现杂交；检测 ６ 份现场镜检班氏微丝蚴阳性的血样 Ｐ Ｃ Ｒ 产物全部阳性，８ 份正常对照均阴性；与微丝蚴模拟血样本和 Ｄ Ｎ Ａ 样本的 Ｐ Ｃ Ｒ 扩增产物杂交时，敏感性分别达１ 条微丝蚴／１００ μｌ和 ２ ｐｇ；检测人工感染的班氏丝虫阳性蚊虫扩增产物全部阳性，正常蚊全部阴性。
In order to search after a sensitive and specific method for filariasis surveillance, a DNA probe/PCR system was established based on the repeated DNA sequence of pWb 12 of W. bancrofti , to detect microfilaria in human blood samples and filarial larvae in mosquitoes. By means of PCR, the W. bancrofti specific DNA fragment of 490 bp was amplified, then the DNA fragment was harvested and labeled with photo biotin as biotin DNA probe. The result showed that the probe hybridized only with DNA of W.bancrofti mf and DNA of mosquitoes infected with W.bancrofti , but not with DNA of B.malayi mf, normal mosquito DNA and human leukocyte DNA. When the PCR products hybridized with the probe, the sensitivity was 1 mf/100 μl blood and 2 pg DNA of W. bancrofti . The 6 microscopically confirmed microfilaria positive blood samples from endemic area were all positive by DNA probe/PCR system, while all of the 8 normal blood samples form non endemic area were negative. Among 10 PCR products of positive mosquitoes artificially infected with W.bancrofti , all of them demonstrated positive by probe, while normal mosquitoes were all negative.
出处
《中国寄生虫病防治杂志》
CSCD
1999年第3期197-199,共3页
Chinese Journal of Parasitic Disease Control
