牛肉中疯牛病特殊风险物质荧光RT-PCR检测方法的建立  

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

作　　者：史喜菊[1] 马贵平[1] 李冰玲[1] 杨金良[1] 王宇[1] 刘旭辉[1] 李炎鑫[1] 刘全国[1] 

机构地区：[1]北京出入境检验检疫局,北京100026

出　　处：《农业生物技术学报》2007年第5期752-756,共5页Journal of Agricultural Biotechnology

基　　金：国家科技奥运专项(No.207001000560716)资助

摘　　要：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)标记物胶原纤维酸性蛋白(GFAP)mRNA的荧光RT-PCR检测技术。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,其它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℃加热处理30min对检测结果无明显影响,中枢神经组织在30℃以上室温可以稳定存放4d,在4℃可以稳定冷藏2周,检测结果仍然为阳性。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、快速方便等优点,适合于在日常检测工作中推广使用。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. 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 ℃ heating treatment for 30 min and long period of storage at over 30 ℃ room temperature （RT） for 4 d and at 4 ℃ for 15 d. 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.

关 键 词：疯牛病 风险物质 荧光RT-PCR 牛肉 

分 类 号：S188[农业科学—农业基础科学]