摘要
从吉林某猪场采集曾接种过猪繁殖与呼吸综合征疫苗的发病猪病料,经RT-PCR初步鉴定为猪繁殖与呼吸综合征病毒后,利用反转录合成cDNA,用针对PRRSV M、N基因片段设计的特异性引物进行扩增及电泳,在紫外凝胶成像系统下可见约为660bp特异性扩增条带。解剖发病仔猪,发现其有HP-PRRSV发病的特征,两耳及鼻端淤血,呈蓝紫色,肺部等内脏器官淤血呈暗红色。病理组织切片显示,病猪有典型的间质性肺炎的特征性病理变化。将其接种于Marc-145细胞后,在培养至第4代时出现典型的细胞病变(CPE),表现为细胞聚集成丛、随后固缩、变圆、脱落;经Reed-Muench法计算得出两PRRSV分离株的病毒滴度分别为10-5.30 TCID50/0.1mL和10-5.53TCID50/0.1mL。用间接免疫荧光试验观察到在接种病料的Marc-145细胞胞浆内出现特异性荧光,而未接种PRRSV的细胞对照则未见到荧光反应。
Visceral organs were collected from pigs at one Jilin farm,which vaccine with porcine reproductive and respiratory syndrome virus（PRRSV）vaccines according to the regular immune program,have shown that infected with PRRSV via RT-PCR identification.Therefore,cDNA were synthesized and the both M and N gene were amplified by PCR using specific primers,followed by electrophoresis and observation using Uv gel imaging system,a 660bp band was seen as expected.The anatomy results showed that HP-PRRSV typical pathogenesis symptoms in PRRSV infected piglets,as indicated with the phenomenon of congestion and been violet in ears and nose,as well as visceral organs filled with blood such as lung.Pathological biopsy showed infected pigs has typical characteristic pathological changes of interstitial pneumonia.The isolated viruses were used to incubate the Marc-145cell line.Typical cytopathic effect（CPE）appeared after 4th generations of passage,which manifested as aggregation of the cells into clumps,followed by pyknosis,round-up and detachment.Titers of the two isolated strains of PRRSV were 10-5.30TCID50/0.1mL and 10-5.53 TCID50/0.1mL respectively as calculated by Reed-Muench method.Indirect immunofluorescence assay showed specific fluorescence only appeared in cells infected with PRRSV but not in control group.
出处
《中国兽医学报》
CAS
CSCD
北大核心
2014年第7期1033-1038,共6页
Chinese Journal of Veterinary Science
基金
吉林省科技厅科技支撑计划重点攻关项目(20130206021NY)
关键词
猪繁殖与呼吸综合征病毒
分离
鉴定
吉林株
生物学特性
porcine reproductive and respiratory syndrome virus
isolation
identification
Jilin strain
biological characteristics
