基于阳离子膜滤芯的环境水体病毒浓缩方法效果评价

摘要:目的　本研究评价和优化基于阳离子膜滤芯的环境水体病毒浓缩方法，为监测环境水体中的病毒提供方法依据。方法　采用基于阳离子膜滤芯 NanoCeram 的膜吸附洗脱法作为初级浓缩手段，结合有机絮凝和超滤法，从大体积初始水量中浓缩目标病毒。预实验评价不同有机絮凝条件对病毒基因拷贝数回收和病毒滴度的影响。结果　随着有机絮凝牛肉浸膏洗脱液 pH 值的上升，回收病毒的基因拷贝数存在下降趋势（β＝41.59，P＜0.01）。采用2种浓缩流程回收4种水体中的目标病毒，成组 t 检验显示两者差异有统计学意义（t＝2.261，P＜0.05）。当调节有机絮凝洗脱液 pH 值为 2.0 时，水源水中脊髓灰质炎病毒的回收率最高达到7.87%。结论　本研究建立的方法可作为今后环境水体常规监测的有效手段。

Evaluation on viral concentration of environmental water using electropositive filter

Abstract: Objective The goal of this study was to evaluate a new method using electropositive filters for the recovery of seeded virus from environmental water. Methods In the study, an electropositive cartridge filter, the NanoCeram filter, was evaluated for its ability to concentrate seeded viruses from large volumes of water. Seeded polioviruses were concentrated using the adsorption- elution procedure, followed by organic flocculation and ultrafiltration for secondary and tertiary concentration. The effects on virus recovery and virus titer of various secondary concentration procedures were investigated through pre-experiment. Results When increasing the pH of beef extract eluate, the recovery of virus genome copies decreased (β=41.59, P<0.01). In order to optimize organic flocculation process, two protocols used for four types of water were evaluated. Seeded virus recovery efficiencies with two protocols were statistically different (t=2.261, P<0.05). The highest virus recovery efficiency was 7.87% for poliovirus from source water, when adjusting the pH to 2.0 for organic flocculation procedure. Conclusion This method is an effective low-cost procedure in a routine laboratory working in water monitoring.