城市污水中诺如病毒不同富集浓缩方法的比较研究

秦萌; 杨楠; 郑宇丹; 刘春源; 陈燕; 李金松

doi:10.3784/jbjc.202505110354

城市污水中诺如病毒不同富集浓缩方法的比较研究

Comparative evaluation of diverse enrichment and concentration techniques for norovirus in urban sewage

摘要

摘要:
目的比较磁珠法、聚乙二醇（PEG）沉淀法和阴离子膜吸附–洗脱法富集浓缩污水中诺如病毒（NoV）的效率，选择合适的城市污水中NoV检测富集浓缩方法。
方法采用磁珠法、PEG沉淀法和阴离子膜吸附–洗脱法对加入梯度稀释的3个不同浓度NoV的污水进行富集浓缩，结合实时荧光定量反转录–聚合酶链式反应（RT-PCR）检测富集浓缩后NoV GⅠ 型和 GⅡ型浓度,比较3种富集方法效果差异。
结果对于NoV GⅠ型，磁珠法、PEG沉淀法和阴离子膜吸附–洗脱法的回收率分别为（30.50±2.04）% ～（34.00±4.49）%、（6.40±2.72）%～（11.58±2.21）%和（3.50±1.55）%～（9.26±2.82）%。磁珠法回收率显著高于PEG沉淀法和阴离子膜吸附–洗脱法（F=85.144～109.443，P＜0.001）。对于NoV GⅡ型，磁珠法、PEG沉淀法和阴离子膜吸附-洗脱法的回收率分别为（30.37±2.60）%～（34.00±4.52）%、（6.40±2.70）%～（10.59±2.58）% 和（3.50±1.51）%～（7.23±1.99）%。磁珠法回收率显著高于PEG沉淀法和阴离子膜吸附–洗脱法（F=71.139～86.526，P＜0.001）。对于NoV GⅠ型和 GⅡ型，阴离子膜吸附–洗脱法和PEG沉淀法回收率差异均无统计学意义。磁珠法、PEG沉淀法和阴离子膜吸附–洗脱法的耗时分别为80 min/24份样本、320 min/24份样本、250 min/24份样本；成本分别为160.00元/份、16.00元/份、18.00元/份。
结论综合考虑3种方法回收率、用时长短和操作简便性，磁珠法富集浓缩污水中NoV的效果最佳。

Abstract:
Objective To compare the efficiency of three virus enrichment and concentration methods, i.e. magnetic bead-based extraction, polyethylene glycol (PEG) precipitation, and anion membrane adsorption-elution for enriching norovirus in wastewater and identify the optimal method for norovirus detection in urban sewage.
Methods Sewage samples spiked with norovirus at three different concentrations (serially diluted) were processed using the three enrichment methods mentioned above. The recovery efficiencies of three enrichment methods for norovirus GⅠ and GⅡ were evaluated by using real-time reverse transcription polymerase chain reaction to detect the virus concentrations.
Results For norovirus GⅠ, the recovery rate ranged from (30.50 ± 2.04)% to (34.00 ± 4.49)% by magnetic bead-based extraction, from (6.40 ± 2.72)% to (11.58 ± 2.21)% by PEG precipitation, and from (3.50 ± 1.55)% to ( 9.26 ± 2.82)% by anion membrane adsorption-elution. The magnetic bead-based extraction demonstrated significantly higher recovery rate compared with both PEG precipitation and anion membrane adsorption-elution (F=85.144−109.443, P＜0.001). Similarly, for norovirus GⅡ, the recovery rate ranged from (30.37 ± 2.60)% to (34.00 ± 4.52)% by magnetic bead-based extraction, from ( 6.40 ± 2.70)% to (10.59 ± 2.58)% by PEG precipitation, and from (3.50 ± 1.51)% to (7.23 ± 1.99)% by anion membrane adsorption-elution, with magnetic beads again outperforming the other methods (F=71.139−86.526, P＜0.001). No significant difference was observed between PEG precipitation and anion membrane adsorption-elution in the recovery rate for either norovirus GⅠ or GⅡ. The processing times for 24 samples were 80 min by magnetic bead-based extraction, 320 min by PEG precipitation and 250 min by anion membrane adsorption-elution, with per-sample costs of 160.00, 16.00, and 18.00 yuan, respectively.
Conclusion Considering recovery efficiency, processing time, and operational simplicity, magnetic bead-based extraction is most effective for norovirus enrichment and concentration in urban sewage.

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