摘要
用实验室－２０℃条件下保存６年的禽成髓细胞性白血病病毒（ＡＭＶ）ＢＡＩ－Ａ株鸡传代血浆毒感染１～３日龄的雏鸡，接毒后１５～４５ｄ陆续发病。取其血浆毒，经差速离心和蔗糖密度梯度离心，获得较纯的ＡＭＶ抗原。用此抗原免疫８～１０周龄ＢＡＬＢ／Ｃ小鼠，将其脾细胞与ＳＰ２／Ｏ瘤细胞进行融合，经ＥＬＩＳＡ筛选、克隆，共获得６株阳性杂交瘤细胞株。分别采用传染性支气管炎病毒（ＩＢＶ）、新城疫病毒（ＮＤＶ）、传染性法氏囊病毒（ＩＢＤＶ）、传染性喉气管炎病毒（ＩＬＴＶ）、减蛋综合征病毒（ＥＤＳ－７６）、及火鸡疱疹病毒（ＨＶＴ）等抗原用间接ＥＬＩＳＡ法对阳性杂交瘤细胞进行特异性鉴定，其中２株证明为稳定分泌特异性抗体的杂交瘤细胞系，而对其它病毒则呈阴性反应。用该２株单抗分别进行临床检测，其结果显示。
One to three-day old susceptible chicks were experimentally infected with the BAI-A stain of Avian Myeloblastosis Virus (AMV) in passaged chicken plasma stored at -20℃ for six years, and the onset of disease was seen 15 to 45 days postinoculation. BALB/C mice 8～10 weeks of age were immunized with AMV antigens purified from AMV nfected chicken plasma by differential and isopycnic centrifugation in a discontinuous sucrose gradient. B lymphocytes from mouse spleen were hybridized with SP2/O myeloma cells, and six hybridoma cell lines finally established after specific antibody detection and cell cloning. Indirect ELISA demonstrated that only two ascitic monoclonal antibodies (McAb) reacted positively with specific AMV antigen and showed no cross reaction with IBV, IBDV, EDSV-76, ILTV and HVT viruses, retaining its specific antibody secreting property. Futhermore, the application of AMV-McAb in diagnosis and eradication of avian leukosis from chicken flocks was discussed.
出处
《华南农业大学学报》
CAS
CSCD
北大核心
1997年第2期82-84,共3页
Journal of South China Agricultural University
关键词
抗原
单克隆抗体
杂交瘤细胞
鸡病
AMV
avian myeloblastosis virus
group specific antigen
monoclonal antibody
hybridoma
