原位PCR及其在兽医分子生物技术中的应用

Study on in-situ PCR Technology and its Application in Veterinary Molecule Biotechnology

原位 PCR是一种将常规 PCR的高效扩增与原位杂交技术结合起来的新方法 ,其类型主要有直接原位 PCR、间接原位 PCR、原位 RT-PCR和原位再生序列复制反应 ,其中间接原位PCR应用最为广泛。原位 PCR技术的基本步骤包括标本处理、蛋白酶消化、原位 PCR扩增和扩增后产物的检测。原位 PCR技术操作比较复杂 ,同时在操作中易出现假阳性结果和假阴性结果。尽管如此 ,原位 PCR技术作为一种特异性、敏感性高的靶序列定位检测技术 ,作为研究基因信息和细胞、染色体等的形态信息的有力工具 。

In-situ PCR technology is a new method which combines the normal high copy PCR with in-situ hybridization. It contains direct in-situ PCR,indirect in-situ PCR,in-situ RT-PCR and in-situ regenerate sequence copy. Indirect in-situ PCR is used widely. The elementary procedure of this technology contains sample disposal, protein enzyme digestion,in-situ PCR and the detection of sequence copied. The procedure of in-situ PCR is complex, and the pseu-positive and pseu-negative results are easy to occur. Even so, as a kind of special and high sensitive technology to detect the place of sequence needed and a good tool to study information of gene and the shape of chrosome of cell,in-situ PCR has good applications and prospect in veterinary molecule biotechnology.