环境水体中肠道病毒的膜吸附-洗脱浓缩方法研究

Study on Membrane Adsorption-Elution Method for Concentration of Enteroviruses from Environmental Waters

在膜吸附-洗脱和洗脱液浓缩相结合的基础上,建立了一种简便实用的水中肠道病毒浓缩方法.通过实时定量RT-PCR检测,比较了不同材料和不同孔径的微孔滤膜对病毒的吸附效果;对膜洗脱方式进行了改进;研究了在洗脱液浓缩过程中,PEG浓度对于病毒回收率的影响.最后确定了最佳的浓缩方法.选择效果好而且来源广泛的0.22μm孔径的混合纤维素酯微孔滤膜,采用磁力搅拌来洗脱滤膜上吸附的病毒;洗脱液浓缩步骤中,PEG最佳质量浓度为130 g/L.系统比较了不同病毒接种量下,方法中各步骤的病毒回收率.对接种已知量的肠道病毒的生活污水、二级处理出水和地表水等样品的试验结果表明,该方法效果稳定,适合不同水样中肠道病毒的浓缩分离.

A simple and efficient method for concentration of enteroviruses from water sample was established based on the membrane adsorption-elution method combined with eluant concentration. By means of real-time RT-PCR, microporous filters with various nominal pores and materials were compared ; the membrane elution method was modified; the effect of PEG on virus recovery was studied; the optimal method was determined at last. In view of the virus recovery and cost, cellulose mixed-ester microporous filter with nominal pore size of 0. 22μm was superior to other filters. Magnetism agitation was used for membrane elution and the optimal final mass concentration of PEG was 130 g/L in eluant concentration step. The recovery of virus was studied in every step in various seeding viruses. The method was tested in surface water, secondary effluent and sewage seeded with viruses. The results showed that the modified method was reliable and suitable for separation and concentration of enteroviruses from various water samples.