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猫后睾吸虫特异性DNA探针的分离与鉴定

Isolation and characterization of Opisthorchis viverrini specific DNA probe.

作者信息

Sermswan R, Mongkolsuk S, Panyim S, Sirisinha S

机构信息

Department of Microbiology, Faculty of Science, Mahidol University, Bangkok, Thailand.

出版信息

Mol Cell Probes. 1991 Dec;5(6):399-407. doi: 10.1016/s0890-8508(05)80011-8.

Abstract

An Opisthorchis viverrini specific repetitive DNA fragment was isolated from the O. viverrini genome. The cloned fragment, herein designated pOV-A6, contained tandemly repeated 334 bp O. viverrini DNA and has the potential to be a highly specific and sensitive DNA probe. Radioactively labelled pOV-A6 could detect as little as 25 pg of its own genomic DNA and five purified eggs by dot-blot hybridization. The presence of 50 O. viverrini eggs could be readily detected in samples containing faecal materials. The probe was highly specific for O. viverrini DNA and did not cross hybridize with DNAs prepared from related and unrelated parasites, enteric bacteria and a number of definitive and intermediate hosts. The probe has potential for screening a large number of samples and could be readily applied in epidemiological studies.

摘要

从猫后睾吸虫基因组中分离出一个猫后睾吸虫特异性重复DNA片段。克隆的片段,在此命名为pOV-A6,包含串联重复的334 bp猫后睾吸虫DNA,有潜力成为一种高度特异且灵敏的DNA探针。放射性标记的pOV-A6通过斑点印迹杂交能够检测低至25 pg其自身基因组DNA以及5个纯化的虫卵。在含有粪便物质的样本中能够轻易检测到50个猫后睾吸虫虫卵。该探针对于猫后睾吸虫DNA具有高度特异性,不会与从相关和不相关寄生虫、肠道细菌以及一些终末宿主和中间宿主制备的DNA发生交叉杂交。该探针有潜力用于筛查大量样本,并且能够轻易应用于流行病学研究。

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