DEVELOPMENT OF A PCR-BASED ASSAY FOR THE DETECTION OF CHIKEN FECES

该子项目是利用NIH/NCRR资助的中心赠款提供的资源的许多研究子项目之一。子项目和研究者（PI）可能从另一个NIH来源获得了主要资金，因此可以在其他CRISP条目中表示。所列机构为中心，不一定是研究者所在机构。水的质量和安全是重大的公共卫生问题。我们的家庭和娱乐场所的水应该是安全的，可以饮用或游泳，并且相对没有污染物，包括粪便污染物。许多人类疾病，如霍乱、伤寒、沙门氏菌病和痢疾，可以通过粪便污染的水传播。本研究的目的是建立一种基于PCR的检测鸡粪便污染的方法，该方法使用来自拟杆菌目的粪便厌氧菌。先前的研究表明，存在于不同物种或具有共同特征的一组物种（例如反刍动物）消化道中的拟杆菌目菌株在遗传上是不同的。已经开发了基于16 S rRNA基因序列差异的检测方法来鉴定人类和反刍动物的粪便污染，但是还没有一种检测方法来区分鸡粪便污染和其他类型的粪便污染。因此，本研究的重点是寻找一种鸡粪便特异性的遗传标记，并建立一种基于PCR的鸡粪便污染检测方法。将使用其他动物的粪便DNA样本检测该试验的专属性。将通过分析含有已知粪便污染物的水样以及测试从当地流域采集的水样来进一步测试该检测方法。最后，将检测检测试剂盒定量不同水平粪便污染的能力和灵敏度限度（上限和下限）。本研究的长期目标是开发和实施快速，特异，灵敏和廉价的分子方法，用于确定水样中粪便污染的来源。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Water quality and safety are major public health issues. The water in our homes and recreational areas is expected to be safe to drink or swim in and to be relatively free of pollutants including fecal pollutants. Many human diseases such as cholera, typhoid, salmonellosis, and dysentery can be transmitted through fecal contamination of water. The goal of this research project is to develop a PCR-based assay for chicken fecal contamination using fecal anaerobic bacteria from the order Bacteroidales. Previous studies have shown that Bacteroidales strains present in the digestive tracts of different species, or in a group of species sharing a common characteristic (e.g. ruminants), are genetically distinct. Assays based on differences in 16S rRNA gene sequences have already been developed for the identification of human and ruminant fecal contamination, but an assay to distinguish chicken fecal contamination from other types of fecal pollution is not yet available. Therefore, the focus of this project is to identify a genetic marker specific for chicken feces and develop a PCR-based assay for chicken fecal contamination. The assay will be tested for specificity using fecal DNA samples from other animals. The assay will be further tested by analysis of water samples containing known fecal contaminants and by testing water samples taken from local watersheds. Finally, the assay will be tested for its ability to quantitate different levels of fecal contamination and for limits of sensitivity (upper and lower). The long-term objective of this research is to develop and implement rapid, specific, sensitive, and inexpensive molecular methods for the determination of the source or sources of fecal contamination in water samples.