猪圆环病毒2型体外滚环复制方法的建立

通过建立猪圆环病毒2型(PCV-2)体外滚环复制(RCA)方法,为PCV-2体外检测探寻更为简便易行的路径。先将西北农林科技大学预防实验室保存分离纯化的PCV-2病料以传统PCR方法扩增出全基因组片段,然后在没有PCR扩增仪的情况下,只用简单的恒温装置,摸索RCA扩增条件,建立体外扩增PCV-2全基因组的方法。最后将2种方法扩增的全基因组片段进行测序,对其相似性进行比对。结果表明,以传统的PCR方法和RCA方法均扩增出1 767bp大小的片段,测序发现两者相似度达到99.9%。可见,成功建立了PCV-2体外滚环复制方法,扩增出病毒的全基因组片段。

Establishment of the Rolling Circle Replication of Porcine Circovirus Type 2 in Vitro

The aim of the study is to establish the rolling circle replication of PCV-2 to explore a more simple path to detect the PCV-2 in vitro.Firstly,the whole genome fragment was amplified from PCV-2 which was saved in Northwest A&F University Prevention Lab by conventional PCR methods.Then,the RCA amplification conditions were determined,which would be used to establish the amplification method of the whole genome fragment of PCV-2 in vitro,the whole process was implemented without PCR amplification.The whole genome fragments amplified by PCR and RCA were sequenced and compared.The results showed that,both of the two methods amplified the 1 767 bp products,shared the similarity of 99.9%.This test successfully established the rolling circle replication method of PCV-2 in vitro,and amplified the whole genome fragment.