用于灵敏检测淡水中反刍动物粪便污染的定量PCR方法及其在高寒岩溶地区的方法评估
Quantitative PCR method for sensitive detection of ruminant fecal pollution in freshwater and evaluation of this method in alpine karstic regions.
作者信息
Reischer Georg H, Kasper David C, Steinborn Ralf, Mach Robert L, Farnleitner Andreas H
机构信息
Institute for Chemical Engineering, Gene Technology Group, Vienna University of Technology, Getreidemarkt 9-166/5, A-1060 Vienna, Austria.
出版信息
Appl Environ Microbiol. 2006 Aug;72(8):5610-4. doi: 10.1128/AEM.00364-06.
Abstract
A quantitative TaqMan minor-groove binder real-time PCR assay was developed for the sensitive detection of a ruminant-specific genetic marker in fecal members of the phylum Bacteroidetes. The qualitative and quantitative detection limits determined were 6 and 20 marker copies per PCR, respectively. Tested ruminant feces contained an average of 4.1 x 10(9) marker equivalents per g, allowing the detection of 1.7 ng of feces per filter in fecal suspensions. The marker was detected in water samples from a karstic catchment area at levels matching a gradient from negligible to considerable ruminant fecal influence (from not detectable to 10(5) marker equivalents per liter).
摘要
开发了一种定量TaqMan小沟结合剂实时PCR检测方法，用于灵敏检测拟杆菌门粪便成员中的反刍动物特异性遗传标记。所确定的定性和定量检测限分别为每个PCR 6个和20个标记拷贝。经检测，反刍动物粪便平均每克含有4.1×10⁹个标记当量，这使得在粪便悬浮液中每过滤器可检测到1.7纳克粪便。在一个岩溶集水区的水样中检测到该标记，其水平与反刍动物粪便影响从可忽略不计到相当大的梯度相匹配（从每升不可检测到10⁵个标记当量）。
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