猪流感病毒分离鉴定

Isolation and Identification of Swine Influenza Virus

用非免疫鸡胚从我国上海、福建、湖北分离到3株猪流感病毒,分别命名为SSH1、SFJ1和SHB1。SIV分离株第5代病毒液对0.7％豚鼠红细胞的血凝活性分别1:2~8、1:2~7和1:2~8,与H_3亚型标准血清血凝抑制价1:2~8、1:2~7和1:2~8,但不能被鸡新城疫阳性血清所抑制。纯化的病毒在电镜下观察,可见到猪流感典型的病毒粒子。病毒液接种于小白鼠,可表现出临床症状,剖杀后可观察到病毒性肺炎。猪体回归试验,可表现出临床症状和病理变化,从肺脏中可分离到病毒并且RT-PCR也可检测到病毒相应的基因片段。从纯化的病毒中提取RNA,进行RT-PCR,可扩增出预期的条带。

Three H_3 subtype strains of Swine influenza virus(SIV), which were named SSH1, SFJ1 and SHB1,were isolated from Shanghai, Fujian and Hubei. These isolates could grow and replicate in chicken embryo, and the hemagglutinin(HA) values were 1:2~8, 1:2~7 and 1:2~8, respectively. The hemagglutinin inhibitor(HI) titers of H^3 subtype antiserum were 1:2~8, 1:2~7 and 1:2~8, respectively, whereas they were negative in HI assays with Newcastle disease antiserum. Under electron micrograph, the virions showed various shapes. The mice infected with allantoic fluid could arise syndromes and pathology; SIV could be isolated again and amplified by RT-PCR from the lung. RNA was extracted from SIV which was purified from infected allantoic fluid, and expected band was observed via RT-PCR.