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DEVELOPMENT OF A NOVEL COX-II MITOCHONDRIAL DNA MARKER FOR RAT FECAL CONTAMINATION.**

Abstract

Rodent feces can contain dangerous human microbial pathogens that are a serious health hazard to the human population. Currently there is no known rapid assay to detect rat fecal contamination. In this study, a rapid PCR-based mitochondrial DNA primer pair was constructed as it enabled the detection of rat fecal pollution in environmental samples. Environmental samples were all tested using end-point PCR after filtration and then extraction of DNA using the DNeasy Powersoil pro kit. These primers were designed to specifically detect rodent feces that belong to the Rattus Sp. taxonomic group. Rodent samples from various counties in Georgia were obtained and used as positive controls. PCR primers for rat COX-II mitochondria DNA were used to specifically detect rat fecal DNA from several rat samples. The primers did not cross react with human, dog, cow, deer, chicken, mice, or squirrel fecal DNA. The rat fecal PCR assay was then successfully used to detect rat fecal pollution in estuarine water samples. The link between prominent disease-causing pathogens and rat fecal pollution in heavily populated areas is cause for concern. This study can then aid in rapid detection of rats, enabling the strategic construction of disease management protocols.

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