RT-PCR检测健康猪扁桃体和流产死胎猪瘟病毒的应用及RFLP分析

Application of RT-PCR detecting hog cholera virus in tonsil of healthy pigs and in samples of abortions and dead born and its RFLP analysis

本研究用反转录＿聚合酶链反应(RT＿PCR)技术对广西部分猪场的健康猪扁桃体材料以及流产胎儿和死胎中的猪瘟病毒(HCV)进行检测。70份健康猪扁桃体材料中阳性11份,阳性率达15 7%;52份流产胎儿及死胎材料中检出阳性9份,阳性率达17 3%。表明广西猪场中有比较严重的健康带毒情况。用限制性片段长度多态性分析法(RFLP)对25份阳性材料、HCLV和石门毒的引物A1/A2的PCR产物进行分析,发现HCLV、石门强毒和广西流行野毒的RT＿PCR扩增产物中均有RsaⅠ的一个识别序列,但HCLV上的识别序列所在位置不同。本研究表明,用引物A1/A2作RT＿PCR检测,并结合RFLP分析技术可把兔化弱毒和流行野毒区分开来,这将对预防猪瘟起到十分重要的作用。

In this study reverse transcription-polymerase chain reaction (RT-PCR) was applied to detect hog cholera virus (HCV) in tonsils of the healthy pigs and in samples of the abortions and the dead born. The results showed that the positive of 70 tonsils of healthy pigs from different places in Guangxi rate was 15.7 %,that 9 of 52 the abortions and the dead born were positive the positive rates was 17.3 %. The result demonstrated that HCV exists in healthy pigs in Guangxi pig farms and this kind of HCV infecti on is serious and has become one of the reasons resulting in breeding disorders. The restrictive fragment length polymorphism (RFLP) was applied to analyze 25 of those PCR products which were synthesized from nonstructural gene P120 of HCV with primers A1/A2. We found that all HCLV, Shimen strain and Guangxi field HCV strains have one recognition site of Rsa I in the PCR products from gene P120. However, HCLV has different locus of recognition site from other strains. This research demonstrated that HCLV could be distinguished from the Guangxi field strains by PCR with primers A1/A2 combining with RFLP technique, the findings would play an important role in preventing and controling Hog Cholera.