禽腺病毒的分离鉴定及LMH细胞适应毒的生物学特性研究

Isolation and Identification of a Fowl Adenovirus and the Biological Characteristics of LMH-adapted Virus Strain

关键词:  禽腺病毒  鸡肝癌细胞  Hexon基因  致病性

摘要:

为对采集自北京平谷某养殖场发病鸡中出现心包积液综合征症状的病料进行病毒分离和鉴定，将病料接种LMH细胞进行病毒分离，并在LMH上连续传代，取C5和C15代细胞毒分别接种3周龄SPF鸡进行致病性试验。根据分离病毒在LMH上的CPE特征、病毒血清中和试验、Hexon基因序列比对分析、SPF鸡致病性回归试验，表明从发病鸡病料中分离的一株病毒为血清4型禽腺病毒；病毒在LMH细胞盲传3代后，就能很好地适应细胞培养，出现禽腺病毒典型的CPE；培养至第10代，病毒在接种后 72h，病毒含量即可达到107.4TCID50/0.1ml的峰值滴度；接种细胞毒的SPF鸡出现4型禽腺病毒导致的典型临床症状和特异性剖检病理变化，两组接种鸡的死亡率分别为100%和70%。本研究分离到的4型禽腺病毒毒株对SPF鸡具有高致死率，但病毒经过细胞连续传代后对鸡的致死率降低。

Abstract:

To isolate and identify the virus from a case of sick chickens with Hydropericardium syndrome. The suspected sample was inoculated on LMH for viral isolation and then passaged for 15 times, and the pathogenicity of C5 and C15 were performed on 3-week SPF Chickens. A serotype 4 fowl adenovirus was identified by cytopathology effect on LMH, serum neutralization, sequence analysis of Hexon and pathogenicity regression test. The virus could adapt to LMH after three blind passages, and LMH infected with isolate strain showed obvious cytopathology effects, the virus titer of the 10th passage was up to 107.4TCID50/0.1ml after 72h inoculated. 3-week SPF Chickens inoculated with C5 and C15 had typical clinical symptoms and specific autopsy pathological changes with a morbidity of 100% and 70%, respectively. The study demonstrated that the serotype 4 fowl adenovirus could cause high mortality to SPF chickens, but after continuous passage on LMH, the mortality rate was reduced.