逆转录-聚合酶链反应在乙型脑炎减毒活疫苗生物反应罐清洁验证中的应用

Reverse transcription polymerase chain reaction apply to detect the residual virus in the bioreactor of Japanese encephalitis virus in the clean Authentication

为了对乙型脑炎减毒活疫苗生物反应罐清洁后乙型脑炎病毒(JEV)检测方法进行探讨,从GenBank中收录的乙型脑炎病毒的E蛋白基因序列设计一对引物,以乙型脑炎减毒株SA14-14-2培养物提取RNA作为模板,进行逆转录和PCR扩增。结果表明乙型脑炎减毒株SA14-14-2扩增出预期的特异性条带,阴性对照没有扩增出任何条带。聚合酶链反应与血吸附试验比较,有灵敏、快速、稳定性的特点,可用于生物反应罐清洁后乙型脑炎残留病毒的检测。

To detect the residual Japanese encephalitis virus in the bioreactor after clean. One paris of primer was designed according to the complete genomic RNA sequence of JEV. RNA was extracted from BHK21 cell inoculated with JEV, As the complete, RNA was amplified by RT-PCR, The results showed that RNA had been amplificated within the specific sequences, it was identical to the expected nucleotide sequences, Non-JEV speciments had not shown specific amplification fragments. RT-PCR is more sensitive, rapid and reliable method in detection residual virus of JEV compared to that hemadsorption test. the RT-PCR is a valuable tool for detection the residual virus in the bioreactor.