Real-time PCR和常规PCR方法快速鉴定肉骨粉中的牛源性成份  

Real-time PCR and conventional PCR detection of bovine materials in meat and bone meal

作　　者：李林[1] 陈继明[1] 王志亮[1] 

机构地区：[1]农业部动物检疫所国家外来动物疾病诊断中心

出　　处：《中国预防兽医学报》2005年第1期50-54,共5页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家自然科学基金 (编号 :30 2 70 981)

摘　　要：疯牛病是一种严重危害动物和人类健康的疾病 ,其病原朊病毒可以通过食物链进行传播 ,因此建立准确的检测牛源性成份的方法非常重要。我们采用常规PCR和Real_timePCR中的TaqMan技术建立了快速鉴定牛源性成份的方法 ,使用Chelex_10 0提取肉骨粉中的DNA ,过程简单可靠仅需要 2 0min左右。对不同含量的牛肉骨粉进行检测 ,常规PCR能够检测到的最低含量是 0 0 0 1% ,而Real_timePCR对相同的样品进行检测所得到的灵敏度更高 ,最高可达到 0 0 0 0 1% ,两种PCR方法的鉴定过程都非常简单快速 ,各需要 2h左右。BSE is a severe disease that threatening human beings and animal health.Prion,the pathogen of BSE,can be transmitted through food chains.So,it is very important to establish an acute method to detet the bovine material from imported meat bone meal (MBM).Here,A TaqMan technology of Real-time PCR and a conventional PCR were developed to detect the bovine material in the MBM.Template DNA of bovine was extracted from MBM using Chelex-100,and this step spent only about 20 minutes.The conventional PCR could detect the 0.001?% of bovine MBM.The sensitivity of the Real-time PCR was even higher,it could detect 0.0001?% of the bovine MBM using the same sample.The processes of two PCR methods were all sensitive,simple and rapid, and spent about two hours respectively.These two methods can be used as the routine assays for bovine material detection in the labs based on their each condition.

关 键 词：牛海绵状脑病 肉骨粉 Real—tinm PCR TAQMAN探针 

分 类 号：S852.51[农业科学—基础兽医学]