摘要
参照Genbank已发表的猪圆环病毒2型的基因序列,取其比较保守的基因片段ORF2,利用引物设计软件Oligo5.0设计两对PCV2型特异的引物,其中第一对引物扩增跨度为ORF2全基因片段,长度为702 bp,第二对引物扩增ORF2中间的一个小片段,跨度大小为494 bp。首先用第一对引物扩增阳性DNA,阳性DNA的浓度从10^(-1)稀释到10^(-11),然后以第一次PCR产物为模板,用第二对引物进行PCR,发现这种套式PCR的方法比用普通PCR灵敏度提高10~2倍。同这种套式PCR方法对广东省市场上出售的几种猪用弱毒疫苗进行猪圆环病毒2型检测,结果发现这几种疫苗全部为阴性,本实验说明目前广东市场上出售的弱毒疫苗在制作过程中没有受到PCV2的污染,解除了广大猪场主的顾虑。
Two pairs of primers, aimed at the conserved segment ORF2, were designed to detect PCV2. The sequences of ORF2 were amplified by the first pair of primers. The amplification was 702 bp. A 494 bp in the ORF2 was obtained by the amplification of the second pair of primers. The second pair primers were able to produce PCR amplification of ORF2. This nested PCR was performed for positive virus and sensitivity was increased by 100 times. This method was applied to detect PCV2 ORF2 segment in 6 swine live virus vaccines. Negative result was observed in the vaccines.
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
2004年第5期328-331,共4页
Chinese Journal of Preventive Veterinary Medicine
