PCR Assay Specific for Chicken Feces.

Cindy R Cisar, Tatsuya Akiyama, Jonathan Hatley, Lori Arney, Nebojsa Kezunovic, Daniel Owen
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Abstract

Bacteroidales are fecal anaerobic bacteria that are common in the digestive systems and feces of warm-blooded animals. Some strains of Bacteroidales have been reported to be host-specific. In this study, Bacteroidales strains from chicken feces were examined for their potential use as indicators of chicken fecal contamination. Bacteroidales 16S rRNA gene sequences from chicken feces were amplified, cloned and sequenced. Phylogenetic analysis was performed using these sequences and published Bacteroidales 16S rRNA gene sequences from human and bovine feces. Primers were designed based on putative chicken feces-specific 16S rRNA gene sequences and the primer pairs were tested for specificity in PCR assays. One set of primers, chBact F1 and chBact R16, specifically amplified DNA from chicken feces in a PCR assay, but did not amplify wild turkey, cat, bovine, or deer fecal DNAs. In addition, DNA from feces contaminated straw-based chicken litter produced a product in the PCR assay. However, DNA from feces contaminated wood shavings-based chicken litter was not amplified. The PCR assay described here may prove a useful tool for the detection of chicken feces and for source tracking in watersheds with fecal contamination.

Abstract Image

Abstract Image

鸡粪便特异性PCR检测。
拟杆菌属是在温血动物的消化系统和粪便中常见的粪便厌氧细菌。据报道,一些拟杆菌属菌株具有宿主特异性。本研究对鸡粪便中的拟杆菌菌株进行了检测,探讨其作为鸡粪便污染指标的潜力。对鸡粪便中拟杆菌16S rRNA基因序列进行扩增、克隆和测序。将这些序列与已发表的人类和牛粪便拟杆菌属16S rRNA基因序列进行系统发育分析。根据推测的鸡粪特异性16S rRNA基因序列设计引物,对引物进行PCR特异性检测。其中一组引物,chBact F1和chBact R16,在PCR检测中特异性扩增鸡粪便中的DNA,但不能扩增野生火鸡、猫、牛或鹿粪便中的DNA。此外,从粪便中提取的DNA在PCR检测中产生了一种产物。然而,从受木屑污染的鸡窝粪便中提取的DNA未被扩增。本文所描述的聚合酶链反应试验可能是一种有用的工具,用于检测鸡粪便和追踪粪便污染流域的来源。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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