牛肉中疯牛病特殊风险物质荧光RT-PCR检测方法的建立

本研究建立了检测牛肉中疯牛病特殊风险物质（主要是中枢神经组织）标记物GFAP mRNA的荧光RT-PCR检测技术。试验结果表明：该技术具有良好的种属特异性和组织特异性，只能从牛源和羊源的中枢神经组织中检测到GFAP，猪源和禽源检测结果为阴性，而且只有脑和脊髓等中枢神经组织产生阳性反应，其他内脏组织以及不同部位牛肉检测结果均为阴性；敏感性检测结果表明，该方法最低检测限达到0.001%以下；稳定性试验结果表明，100°C 加热处理30min对检测结果无明显影响，中枢神经组织在30°C以上室温可以稳定存放4天， 在4°C可以稳定冷藏2周，检测结果仍然为阳性。该结果显示，所建立的荧光RT-PCR技术用于牛肉中疯牛病特殊风险物质的检测具有特异性强、敏感性高、稳定性好及快速方便等优点，适合于在日常检测工作中推广使用。

Detection of Bovine Central Nervous System Tissue as Bovine Spongiform Encephalopathy Specified Risk Material in Beef by Real Time RT-PCR

A real time RT-PCR detection of Bovine central nervous system tissue （CNS）as bovine spongiform encephalopathy（BSE）specified risk material （SRM） in beef and beef products based on glial fibrillary acidic protein (GFAP) mRNA was reported in this study. The results showed that the developed method allowed the detection of CNS tissues from bovine and ovine origins, but not from porcine and avian origins, and that the lowest detection limit was below 0.001% bovine brain homogenate, and the GFAP mRNA signal detection was not affected by 100°C heating treatment for 30min and long period of storage at over 30°C room temperature (RT) for 4 days and at 4°C for 15 days. It is concluded that real time RT-PCR based on GFAP mRNA can serve as a sensitive and specific test in routine inspection and quarantine detection for illegal use of bovine CNS tissues in beef and beef products.