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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。
Objective To establish an indirect immunofluorescence assay (IFA) for the detection of SAdV-1 in serum and provide a reference for the detection of SAdV-1 infection in experimental monkeys. Methods BSC-1 cells were infected with SAdV-1 virus. When lesions occurred in 50% of cells, trypsin digestion was performed on 24-well coated glass slides with 20 μL of 1 × 107 cells / mL and fixed with acetone. The optimal working conditions of goat anti-monkey secondary antibody were determined by the titration of the prepared antigenic tablets. IFA specificity, sensitivity, reproducibility validation and preliminary detection of monkey serum samples. Results The indirect immunofluorescence assay of SAdV-1 antibody was established. Among 21 monkey serum samples collected, 9 cases were positive for SAdV-1 antibody and 12 cases were negative for serum test. Conclusion The method has good specificity and stability, which can be used as a reliable method for detecting SAdV-1.