BDV核蛋白FQ RT-PCR试剂盒的评价与应用

Verification and Practical Application of BDV Nucleoprotein FQ RT-PCR Kit

摘要:

为评价博尔纳病病毒(Borna disease virus, BDV)核蛋白荧光定量PCR(FQ RT-PCR)试剂盒的各项指标, 比较分子信标探针相对普通探针的优势, 并了解其实际检测效果, 本课题组使用BDV OL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。实验结果显示: 试剂盒可以检测出的病毒RNA最低浓度为2.5×101, 相当于1个病毒拷贝数, 无非特异检出; 不同批次的试剂盒的检测结果变异系数小于0.7; 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4.2% (猪)和1.5% (马)。可见该试剂盒重复性和稳定性均好; 敏感性、特异性优于普通探针试剂盒, 是BDV基础研究、流行病学调查和临床检测的良好工具。

Abstract:

In order to verify the Fluorescent-Quantitation PCR kit specific for Borna disease virus (BDV) nucleoprotein, compare molecular beacon with common probe and study its practical application, the sensitivity, distinctness, repeatability and stability were tested on BDV and non-BDV infected cells, as well as non-infected controls. Human and animal samples were also tested by the kit. The lowest quantity of virus RNA detected by the kit was 2.5×101, which corresponded to 1 virus copy. No false positive result was found. The coefficient of variation (CV) of different batches was less than 0.7, and the CV between normal kits and the kits treated by accelerating decomposition was less than 2. The morbidities of human and animal detected by the kits were 3.6% (human), 4.2% (swine) and 1.5% (horse). The sensitivity and distinctness of this kit are better than common probe kit. All of the results indicate that this kit is an efficient tool in fundamental research, clinical detection and epidemiological investigation of BDV.