摘要
为获得免疫原性好的禽腺病毒4型毒株,试验将滨州某养鸡场疑似腺病毒感染的鸡肝脏组织处理后,接种鸡肝癌(LMH)细胞,取细胞培养物提取DNA,进行PCR鉴定及测序分析,同时进行动物回归试验和免疫原性研究。结果表明:试验成功分离到1株禽腺病毒,分离毒株接种LMH细胞,第1代即可产生明显的细胞病变。分离毒株在LMH细胞上第8代的细胞培养物PCR鉴定结果为阳性,测序结果与NCBI序列比对分析显示,分离毒株与禽腺病毒4型核苷酸相似性最高,达100%,表明分离毒株为禽腺病毒4型。该毒株能致21日龄SPF鸡死亡,死亡率达100%,且分离毒株攻毒后能够完全复制出与临床自然感染鸡相同的症状。用分离毒株制备的3个批次油苗攻毒保护率均为100%,血清琼扩效价几何平均值分别为1∶4.6,1∶4.9,1∶4.6。说明分离得到的病毒为Ⅰ群禽腺病毒4型,该分离株具有很好的免疫原性。
In order to obtain a good immunogenic strain of Fowl aviadenovirus type 4,the experiment processed chicken liver tissue suspected of adenovirus infection in a chicken farm in Binzhou,and then it was inoculated into chicken liver cancer cells(LMH),and the cell culture samples were identified by PCR and sequencing analysis,while animal regression test and immunogenicity studies were conducted.The results showed that one strain of Fowl aviadenovirus was successfully isolated in the experiment,and the isolated strain was inoculated with LMH cells,and the first(F1)generation could produce obvious cytopathic changes.The PCR identification results of the cell culture of the 8 generation of the isolated strain on LMH cells was positive;the sequencing result and NCBI sequence comparison analysis showed that the isolated strain had the highest nucleotide similarity with Fowl aviadenovirus type 4,reaching 100%,which suggested that the isolated strain was Fowl aviadenovirus type 4.This strain could cause the death of 21-day-old SPF chickens with a mortality rate of 100%,and the isolated strain could completely replicate the same symptoms as clinically infected chickens.The three batches of oil seedlings prepared with the isolated strains had a protection rate of 100%,and the geometric mean values of the agar expansion antibodies were 1∶4.6,1∶4.9,and 1∶4.6,respectively.The results suggested that the isolated virus was GroupⅠof Fowl aviadenovirus type 4,and the isolated strain had good immunogenicity.
作者
魏凤
张文通
苗立中
张培培
祖立闯
李玲
王金良
唐娜
沈志强
WEI Feng;ZHANG Wentong;MIAO Lizhong;ZHANG Peipei;ZU Lichuang;LI Ling;WANG Jinliang;TANG Na;SHEN Zhiqiang(Shandong Binzhou Animal Science and Veterinary Medicine Academy,Binzhou 256600,China;Shandong Lvdu Biological Technology Co.,Ltd.,Binzhou 256600,China)
出处
《黑龙江畜牧兽医》
CAS
北大核心
2021年第8期88-91,153,共5页
Heilongjiang Animal Science And veterinary Medicine
基金
山东省现代农业产业技术体系家禽创新团队计划资助项目(SDAIT-11-16)
2017山东省外专双百计划项目
山东省自然科学基金重点项目(ZR2020KC006)。
关键词
禽腺病毒
分离鉴定
免疫原性
细胞病变
SPF鸡
Fowl aviadenovirus
isolation and identification
immunogenicity
cytopathic effect
SPF chicken
