Faculty of Veterinary Science, The University of Melbourne, Parkville, Victoria 3010, Australia.
Mol Cell Probes. 2013 Jun-Aug;27(3-4):153-7. doi: 10.1016/j.mcp.2013.03.002. Epub 2013 Mar 21.
The specific diagnosis of gastrointestinal parasite infections in livestock is central to their control. PCR assays have been developed for routine diagnosis and to overcome limitations of classical methods. Central to the performance of such assays is the effective isolation of the nucleic acids from samples and the elimination of components that are inhibitory to PCR. Here, we directly compared two techniques for the isolation of DNA from strongylid nematode eggs from faecal samples from sheep, and assessed their performance in relation to the sensitivity and specificity of PCR, time required for DNA isolation and ease of use. The results showed differences in the performance of the two isolation techniques, subsequently effecting the PCR results. The main differences related to the time required for DNA isolation, and the elimination of inhibitory substances from the DNA isolated by one technique but not the other.
对家畜胃肠道寄生虫感染的具体诊断是其控制的关键。聚合酶链反应（PCR）检测方法已被开发出来，用于常规诊断并克服经典方法的局限性。此类检测方法的关键是从样品中有效分离核酸，并消除对 PCR 有抑制作用的成分。在这里，我们直接比较了两种从绵羊粪便样本中的强旋线虫卵中分离 DNA 的技术，并评估了它们与 PCR 的灵敏度和特异性、DNA 分离所需时间以及使用方便性的关系。结果表明，两种分离技术的性能存在差异，随后影响了 PCR 结果。主要差异与 DNA 分离所需的时间以及一种技术从 DNA 中消除抑制物质而另一种技术不能消除有关。
