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禽成髓细胞性白血病病毒的繁殖及其杂交瘤细胞株的建立
PROPAGATION OF AVIAN MYELOBLASTOSIS VIRUS AND ESTABLISHMENT OF HYBRIDOMA CELL LINES

作  者: ; ;

机构地区: 华南农业大学动物医学系

出  处: 《华南农业大学学报》 1997年第2期82-84,共3页

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.

关 键 词: 抗原 单克隆抗体 杂交瘤细胞 鸡病

领  域: [农业科学] [农业科学] [农业科学]

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