猴腺病毒Ⅰ型间接免疫荧光检测方法的建立与初步应用*

摘要: 目的建立检测血清中猴腺病毒Ⅰ型( SAdV-1) 抗体的间接免疫荧光方法( IFA) ，为检测实验猴群中SAdV-1 的感染情况提供参考依据。方法用SAdV-1 病毒感染BSC-1 细胞，待50% 细胞出现病变时，用胰蛋白酶消化以 20μL 1 × 107 个/mL 浓度的细胞液滴到24 孔镀膜玻片上，丙酮固定。利用制备的抗原片通过浓度滴定确定猴血清、 羊抗猴二抗最佳工作条件。对IFA 进行特异性、敏感性、重复性验证并初步检测猴血清样本。结果建立了SAdV- 1 抗体的间接免疫荧光检测方法。在采集的21 份猴血清样本中，检出SAdV-1 抗体阳性9 例，12 例血清检测为阴 性。结论方法具有良好的特异性和稳定性，可作为SAdV-1 检测的可靠方法。

Establishment and Application of Assay for the Detection of SAdV-1 by Immunofluorescence Assay

Objective Establishing the method of detecting the simian adenovirus typeⅠ antibody with immunofluorescence assay(IFA),to provide reference in the detection of SAdV infections in monkeys.Methods Selected SAdV sensitive cell BSC-1,digesting with trypsin while 50% CPE appear,titrate the cells 20μL with the concentration of 1×107/mL on the slide and fix with acetone.We determine the optimal working concentration by density gradient titration.Making the repetitive verification for testing specificity,sensibility and repeatability of IFA method.Detect 21serum with antigen slide in IFA.Results We established the immunofluorescence assay to test SAdV-1,9 serum was positive and 12 serum was negative.Conclusions This method can test SAdV-1 with high specificity and stability.