多聚酶链反应检测狂犬病毒的研究

Studies on PCR for Safety-detection of Rabies Virus in Samples

应用一对狂犬病毒特异性引物,经逆转录和循环的扩增,从狂犬病毒感染的细胞及加热灭活的这种样本中,都扩增出了与预期相符的带,表明结合热灭活技术可用作狂犬病毒的安全,敏感,特异,快速检测手段。

Polymerase Chain Reaction (PCR) combined with Heat-inactivation is adopted to detect of rabies virus. Using a pair of primers specific for rabies virus, reverse transcription and PCR were performed. The amplified products were visualized after electrophoresis on 2% agarose gel, stained with ethidium bromide and exposed to ultroviolet light. It is shown that:(1) A DNA fragment corresponding in size to the distance between the two primers (329 bp) could be amplified from the nucleic acid extracted from the rabies virus sample (BHK-21 cell infected with Rabies virus Flury strain) but not from controls (BHK-21 cell, HSV, VSV); (2)The same DNA band could be amplified from the rabies virus samples inactivated by heating(60℃ 30 min, 75℃ 30 min, 100℃ 2 min, 100℃ 5 min). According to this result, it is suggested that PCR combined with heat-inactivation techniques could be used as a safe, rapid, specific and sensitive method for detection of rabies virus.