向水样中接种已知量的脊髓灰质炎病毒,然后利用微孔滤膜吸附-洗脱法、滑石粉-硅藻土吸附层吸附-洗脱法、化学絮凝沉淀法和滑石粉-硅藻土絮凝沉淀法等4种不同的浓缩方法对水样中的病毒进行浓缩,并采用实时定量RT—PCR技术对各方法浓缩后样品中的病毒进行扩增和检测。综合考虑病毒回收率和浓缩方法的可靠性与简便性等因素,确定微孔滤膜吸附-洗脱法为最优的浓缩方案。此外,还比较了三种洗脱液和两种二次浓缩方法的效果,并对各步骤的回收率进行了分析。
The four concentration methods including microporous filter adsorption - elution method, talcum powder - diatomaceous earth layer adsorption - elution method, chemical flocculation precipitation method and talcum powder - diatomaceous earth flocculation precipitation method were compared by seeding known concentrations of polioviruses into water samples. The viruses from concentrated samples were detected by real-time RT -PCR. In view of the virus recovery, the reliability and simpleness of the methods, the microporous filter adsorption - elution method was chosen as the optimal concentration method. In addition, the effect of three kinds of eluants, and organic flocculation and PEG - NaCl methods in secondary concentration was compared. The recovery of virus in every step was analyzed.