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病毒病是严重危害养猪业的重要疫病,秦岭山中存在大量野猪,其携带猪源病毒的情况尚不明确。本研究对1只从秦岭山中救治的野生仔猪携带的几种猪重要病毒进行了检测。分别采集了野猪的血液、口腔拭子、眼拭子、鼻拭子及肛门拭子样品,通过PCR或RT-PCR方法分别检测了猪瘟病毒(CSFV)、猪繁殖与呼吸综合征病毒(PRRSV)、猪细小病毒(PPV)、伪狂犬病病毒(PRV)、猪圆环病毒1型(PCV1)和2型(PCV2)核酸,对检测阳性的病毒进行了分离。克隆了PCV2分离株的全基因序列,进行了全基因序列分析。结果显示,在所有类型样品中均未检出CSFV、PRRSV和PPV核酸;在血液中均检测到PCV1、PCV2和PRV核酸;在肛门拭子和口腔拭子中检测到PCV2核酸,口腔拭子中还检测到PCV1核酸;眼拭子和鼻拭子中未检出目标病毒核酸。将血液处理后接种PK15细胞进行病毒分离,细胞盲传8代后,可观察到细胞出现CPE,收集细胞后通过PCR可以检测到PCV1、PCV2和PRV核酸阳性。PCV2分离株的全基因序列长度1 767bp,序列分析表明该分离株与从发现该野猪地区养猪场分离的9个PCV2分离株的全基因同源性在99.3%以上,与GenBank上登录的PCV2陕西株的全基因同源性为96.0%~99.6%,从全基因的同源性分析,可以推测该PCV2野猪分离株极有可能来自家养猪。本研究初步发现,秦岭地区的野猪携带有来自家猪的重要病毒,在该地区家猪传染性病毒病的防控中应考虑通过野猪携带和扩散病毒的问题。
Virus disease is an important disease that seriously damages the pig industry. There are a large number of wild boar in the Qinling Mountains, and the situation of carrying the swine virus is not clear. In this study, we detected several important swine viruses carried by wild piglets treated in the Qinling Mountains. Swine blood samples, oral swabs, eye swabs, nasal swabs and anal swabs were collected and tested for CSFV, PRRSV ), Porcine parvovirus (PPV), pseudorabies virus (PRV), porcine circovirus type 1 (PCV1) and type 2 (PCV2) nucleic acids. The whole genome sequence of PCV2 isolate was cloned and the whole genome sequence was analyzed. The results showed that CSFV, PRRSV and PPV nucleic acids were not detected in all types of samples; PCV1, PCV2 and PRV nucleic acids were detected in blood; PCV2 nucleic acids were detected in both anal swabs and buccal swabs PCV1 nucleic acid was also detected; no target viral nucleic acid was detected in eye swabs and nasal swabs. PK15 cells were inoculated and treated with blood for virus isolation. After 8 generations of cell blind passage, CPE was observed in cells and PCV1, PCV2 and PRV were detected by PCR after cells were collected. The complete genome of PCV2 was 1 767bp in length. The sequence analysis showed that the homology between the isolate and the nine PCV2 isolates from pig farms found in the wild boar was over 99.3%, which was consistent with the PCV2 The homology of whole genome of Shaanxi strain was 96.0% ~ 99.6%. According to the homology analysis of whole genome, it can be inferred that the PCV2 wild boar isolate is most likely from domestic pigs. In this study, we found that wild boars in Qinling area carry important viruses from domestic pigs, and the problem of carrying and spreading virus by wild boar should be considered in the prevention and control of domestic pig infectious virus diseases in this area.