鸡病毒性关节炎病毒的分离与鉴定

ISOLATION AND IDENTIFICATION OF VIRUS OF VIRAL ARTHRITIS IN CHICKENS

从疑似鸡病毒性关节炎的自然病鸡中,分离到一株病毒(VAV-1)。将VAV-1接种鸡胚卵黄囊,可使鸡胚死亡,胚体出血呈紫红色。该病毒可在鸡胚成纤维(CEF)、幼鸡肾(CK)、绿猴肾(Vero)和幼仓鼠肾(BHK-21)等细胞上增殖,引起细胞病变(CPE)。幼鸡人工感染VAV-1后,可从发病鸡分离到相似病毒。病毒抵抗乙醚、氯仿、pH3、3％甲醛溶液和胰蛋白酶,60℃6小时病毒未完全灭活。为双链RNA病毒.电镜观察,病毒颗粒呈球形或近似六角形,有双层衣壳,外径63～75nm。琼脂凝胶沉淀(AGP)试验,出现禽呼肠孤病毒(ARV)特异性交叉反应,证明分离病毒VAV-1株为ARV。

The sample materials were taken from the fibulo-metatarsal joint fluid and the edematous tendon of the naturally occurring cases that were suspected to be chicken viral arthritis. They were inoculated into the yolk sac of chick embryos resulting in the isolation of a strain VAV-1. This strain could be passaged by inoculation into the yolk sac,choroallantoic membrane or allantoic cavity, causing deaths of the infected chick embryos within 3-5 days. The embryo proper showed hemorrhagic purpura, visceral hemorrhage and liver necrosis. The virus could multiply on chick embryo fibro-blast, chick kidney, Vero, and BHK-21 cells causing syncytial pathologic change ( CPE ) with the appearance of cytoplasmic inclusion bodies. When chicks artificially infected with the virus showed the same symptoms and the same virus could be recovered. The virus could resist ether, chloroform, pH3, 3% formalin and trypsin, but could not be completely killed at 60℃ temperature for 6 hours. It could be inhibited by 100 μg/ml BUDR. The infected cells when stained by acridine orange showed apple green color reaction. Under electron microscope the yirions were either spherical or hexogonal with double layers of capsid and 63-75nm in diameter. No agglutination was observed when mixed with the red blood cells of chicken, goose, duck, human O type,rabbit, mice or guinea pig. Agar gel precipitation revealed that VAV-1 showed specific reaction with convalescent sera as well as standard serum against avian reovirus. It thus proves that VAV-1 strain belongs to the avian reovirus.