Application of Genetic Testing on Animal Health Administration and Food Industry

• Main Authors: TUNG, HSIANG-YUN, 童湘芸
• Other Authors: TSAI, MING-SHIUN
• Format: Others
• Language: zh-TW
• Published: 2017
• Online Access: http://ndltd.ncl.edu.tw/handle/31992759784019947259

博士 === 大葉大學 === 生物產業科技學系 === 105 === Genetic testing technology, a rapid development of science and technology, is no longer a theory in the textbooks, but can be applied to our daily life to find out the truths.It is developed using gene specificities to detect whether samples contain the expected genes, or to verify whether containing the gene sequence variations or special genetic modifications. Nowadays, genetic testing technology has been widely used in examining human genetic diseases, food contents, animal health monitoring and species identification. Through continuously researching and validation, quick and accurate genetic testing technologies will provide more information for users to obtain the advantages of early and immediately diagnosis.In the study, two genetic testing platforms were evaluated for their feasibilities in industrial applications, including monitoring specific pathogen free (SPF) animals, and rapid on-site detection for commercially soy products with genetically modified organisms (GMOs). In the first study, to increase sensitivity and save diagnosis time for high-throughput processes, multiplex polymerase chain reaction (mPCR) and DNA biochip techniques were combined to develop a multi-pathogen diagnostic method.Thirteen primer sets were designed for multiplex PCR to detect genes from twelve targeted bacterial and viral pathogens. Using this method, a total of five clinical samples were tested, and the result showed that not only single infection but also co-infection by multi-pathogens can be detected. The results demonstrated that multiplex PCR coupled with a DNA bio-chip was an efficient method to detect multi-pathogens in a reaction. This platform may be a useful tool for quarantine services and disease prevention in animal facilities in the future.Another study for GMO detection method, insulated isothermal PCR (iiPCR) method was evaluated for rapid on-site detection of Roundup Ready Soybean (RRS;event GTS40-3-2) in food. The results showed that the detection endpoint 100% of the RRS iiPCR was found at 0.1% RRS, equivalent to that of a reference real-time PCR (rtPCR). Analysis of nucleic acids of soybean-based processed food products indicated 95% agreement between the iiPCR and rtPCR for RRS detection. Testing soybean milk and tofu samples by using simple pre-treatment methods, the agreements were 80% and 90% between these two methods. Replicate tests of all discrepant samples implied that these samples had trace amounts of RRS, suggesting that the iiPCR system was probably more sensitive than the rtPCR method. With the conveniences and accuracy of iiPCR, it can be a useful point-of-need tool for GMOs on-site detection.