RT-PCR一步法检测甲型肝炎活病毒方法的建立

Establishment of One-step RT-PCR Detection Method of Live Hepatitis A Virus

建立灵敏、特异、快速检测甲型肝炎活病毒(HAV)的方法。提取HAV总RNA,选用HAV高保守区为目标区,设计合成一对引物,通过RT-PCR反应扩增其核酸,用琼脂糖凝胶电泳法检测其扩增产物,紫外灯下观察,约200bp处为目标片段,根据观察结果,计算HAV滴度。RT-PCR一步法快速、灵敏、重复性好、特异性强,可用于甲型肝炎活病毒检测。

A detection method with sensitive, specific, and rapid for live hepatitis A virus （HAV） was established. Total RNA was extracted from HAV, and select its high conservative region as goal region, and design a double primers to amplify its nucleotide sequences by RT-PCR, the amplified product was detected with agarose gel dectrophoresis, and the goal fragment was observed at 200 bp under UV lamp, then titer of HAV was counted. The results showed that this method has high specificity as well rapidness, sensitivity and good repetition. Therefore, RT-PCR method may be used to detect live HAV.