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人及动物粪便样本中主要厌氧菌的PCR检测与定量分析

PCR detection and quantitation of predominant anaerobic bacteria in human and animal fecal samples.

作者信息

Wang R F, Cao W W, Cerniglia C E

机构信息

National Center for Toxicological Research, Food and Drug Administration, Jefferson, Arkansas 72079, USA.

出版信息

Appl Environ Microbiol. 1996 Apr;62(4):1242-7. doi: 10.1128/aem.62.4.1242-1247.1996.

Abstract

PCR procedures based on 16S rRNA gene sequences specific for 12 anaerobic bacteria that predominate in the human intestinal tract were developed and used for quantitative detection of these species in human (adult and baby) feces and animal (rat, mouse, cat, dog, monkey, and rabbit) feces. Fusobacterium prausnitzii, Peptostreptococcus productus, and Clostridium clostridiiforme had high PCR titers (the maximum dilutions for positive PCR results ranged from 10(-3) to 10(-8)) in all of the human and animal fecal samples tested. Bacteroides thetaiotaomicron, Bacteroides vulgatus, and Eubacterium limosum also showed higher PCR titers (10(-2) to 10(-6)) in adult human feces. The other bacteria tested, including Escherichia coli, Bifidobacterium adolescentis, Bifidobacterium longum, Lactobacillus acidophilus, Eubacterium biforme, and Bacteroides distasonis, were either at low PCR titers (less than 10(-2)) or not detected by PCR. The reported PCR procedure including the fecal sample preparation method is simplified and rapid and eliminates the DNA isolation steps.

摘要

基于16S rRNA基因序列,开发了针对在人类肠道中占主导地位的12种厌氧细菌的PCR检测方法,并用于定量检测人类(成人和婴儿)粪便以及动物(大鼠、小鼠、猫、狗、猴子和兔子)粪便中的这些菌种。在所有测试的人类和动物粪便样本中,普氏栖粪杆菌、产物消化链球菌和梭状梭杆菌属的梭状芽孢杆菌具有较高的PCR滴度(阳性PCR结果的最大稀释度范围为10^(-3)至10^(-8))。多形拟杆菌、普通拟杆菌和黏液真杆菌在成人粪便中也显示出较高的PCR滴度(10^(-2)至10^(-6))。测试的其他细菌,包括大肠杆菌、青春双歧杆菌、长双歧杆菌、嗜酸乳杆菌、双形真杆菌和狄氏拟杆菌,要么PCR滴度较低(小于10^(-2)),要么未通过PCR检测到。所报道的PCR检测方法包括粪便样本制备方法,操作简便、快速,且省去了DNA提取步骤。

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