H1亚型猪流感病毒的分离鉴定

Isolation and Identification of H1 Subtype Swine Influenza Virus

从山东、浙江、湖南、广西等地区各猪场采集30份疑似猪流感病猪的病料,通过鸡胚培养、血凝及血凝抑制试验、电镜观察、RT-PCR和测序分析进行了分离鉴定。结果从山东和湖南两地分离到4株有血凝活性的病毒,其中山东的2株病毒与新城疫病毒阳性血清的血凝抑制试验为阳性,通过电镜观察湖南的2株病毒具有猪流感病毒的特征,且与抗H1亚型猪流感病毒阳性血清的血凝抑制试验为阳性;根据H1亚型猪流感病毒HA基因设计引物,利用RT-PCR扩增出543bp片段,经序列分析证实该病毒为H1亚型猪流感病毒。

30 samples were collected from Shandong, Zhejiang, Hunan and Guangxi. The isolated virus were identified by chicken embryo culture, hemagglutination test, hemagglutination inhibition test, electron microscope observation, RT- PCR and sequencing. The results showed that 4 strains virus have hemagglutination activity from Shandong and Hunan. The HI test against new castle disease virus of 2 strains virus from Hunan were positive and the others were negative. By electron microscope observation, the other 2 strains virus have SIV ＇ s feature from Hunan, and the HI test against H1 subtype swine influenza virus were positive. Then a pair of primer was designed according to HA gene of swine influenza virus published in the GenBank. About 543bp fragment can be amplified by RT-PCR. The results confirmed that the virus was H1 subtype swine influenza virus.