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目的评价博尔纳病病毒(Borna disease virus,BDV)实时荧光定量PCR(FQ RT-PCR)试剂盒的各项指标,并了解其实际应用效果。方法使用BDVOL持续感染细胞株、非BDV病毒序列转染的OL细胞、正常的OL细胞,对BDV RT-PCR试剂盒的敏感性、特异性、重复性和稳定性进行评估,同时检测部分临床病人和动物外周血液RNA。结果试剂盒可以检测出的病毒RNA最低浓度为102,相当于1.5个病毒拷贝数。特异性好,无非特异检出。不同批次的试剂盒的检测结果变异系数接近1。加速破坏的试剂盒和正常试剂盒检测结果之间变异系数在2以内。对临床病人检测阳性率为3.6%,对动物检测阳性率为4.4%。结论试剂盒敏感性、特异性、重复性和稳定性均佳,是BDV基础研究、流行病学调查、临床检测的良好工具。
Objective To evaluate the indicators of Borna disease virus (BDV) real-time fluorescence quantitative PCR (FQ RT-PCR) kit and to understand its practical application. Methods The sensitivity, specificity, reproducibility and stability of the BDV RT-PCR kit were evaluated using continuous infection of BDVOL cells, OL cells transfected with non-BDV virus sequences and normal OL cells. Meanwhile, some clinical patients And animal peripheral blood RNA. Results The lowest detectable concentration of viral RNA in the kit was 102, equivalent to 1.5 viral copies. Good specificity, no non-specific detection. Different batches of test results of the coefficient of variation close to 1. The coefficient of variation (CV) between accelerated test and normal test is within 2. The positive rate was 3.6% for clinical patients and 4.4% for animal tests. Conclusion The kit has good sensitivity, specificity, repeatability and stability and is a good tool for basic research, epidemiological investigation and clinical examination of BDV.