一株H3N2亚型猪流感病毒的分离鉴定

为了给猪流感疫苗的研制做准备,试验将从某猪场现地采集疑似猪流感发病猪的鼻拭子样品,经过处理后接种至10~11日龄SPF鸡胚进行盲传,分离到1株具有血凝性的病毒。通过血清学、分子生物学鉴定、电镜观察、动物回归试验等方法对该病毒进行鉴定。结果发现:该病毒可与猪流感病毒H3亚型阳性血清特异性结合;通过分子生物学检测,该分离株出现猪流感病毒H3亚型和N2亚型目的片段;将该分离株液置于电镜下观察可见直径为80~120 nm且具有囊膜和纤突的病毒粒子,符合猪流感病毒粒子形态特征;用纯化后的病毒攻击4~6周龄阴性仔猪,攻毒组仔猪发病率可达80%。由此可见分离获得的病毒为H3N2亚型猪流感病毒。

Isolation and Identification of a H3N2 Subtype Swine Influenza Virus

In order to prepare for the development of swine influenza vaccine,nasal swab samples from pigs suspected of having swine influenza were collected at a pig farm.After treatment,10-11-day-old SPF chicken embryos were inoculated for blind transmission,and a strain of hemagglutinating virus was isolated.The virus was identified by serology,molecular biological identification,electron microscope observation and animal regression test.The results showed that:the virus could specifically bind to the H3 subtype positive serum.Molecular biological detection demonstrated that H3 subtype and N2 subtype target fragments appeared in the isolated strains.The virus particles 80-120 nm in diameter with capsule membrane and fibrillae were observed from the isolated strains liquid under the electron microscope,which is consistent with the morphological characteristics of swine influenza virus particles.When the purified virus was used to attack the negative piglets aged from four to six weeks,the morbidity of piglets in the challenge group could reach 80%.Thus,the isolated virus was H3N2 subtype swine influenza virus.