生活饮用水中五种病毒富集方法的比较  

Comparison of five virus enrichment methods for drinking water

作　　者：谭梦迪 高志勇[2] 赵佳琛[2] 严寒秋[2] 李伟红[2] 张代涛[2] 王全意[2] 石伟先[2] Tan Mengdi;Gao Zhiyong;Zhao Jiachen;Yan Hanqiu;Li Weihong;Zhang Daitao;Wang Quanyi;Shi Weixian(School of Public Health,China Medical University,Shenyang 110122,China;Institute for the Control of Infectious and Endemic Diseases,Beijing Center for Disease Prevention and Control,Beijing 100013,China)

机构地区：[1]中国医科大学公共卫生学院,沈阳110122 [2]北京市疾病预防控制中心传染病与地方病控制所,北京100013

出　　处：《中华实验和临床病毒学杂志》2025年第1期102-108,共7页Chinese Journal of Experimental and Clinical Virology

基　　金：北京市高层次公共卫生技术人才建设项目(领军人才-01-02,学科带头人-02-07)。

摘　　要：目的比较超滤法、聚乙二醇(polyethylene glycol,PEG)沉淀法、铝盐沉淀法、阴离子膜吸附—洗脱法对生活饮用水中病毒的富集效果差异。方法以噬菌体MS2为目标病毒,制备3种不同浓度的生活饮用水样本,采用超滤法1、超滤法2、PEG沉淀法、铝盐沉淀法、阴离子膜吸附—洗脱法分别对样本进行富集,使用实时荧光定量逆转录聚合酶链式反应(quantitative reverse transcription-polymerase chain reaction,RT-qPCR)法对浓缩前后样本中MS2核酸进行定量检测,比较5种方法对高、中、低浓度样本中的MS2回收率。结果对单个富集方法在不同浓度水样中的MS2富集回收率进行比较,超滤法1、PEG沉淀法、铝盐沉淀法、膜吸附—洗脱法均不受到样本浓度的影响,这4种方法在3种浓度样本中的回收率差异均不具有统计学意义(均P>0.05)。5种富集方法在各浓度样本中的MS2富集回收率均具有统计学差异(均P<0.05),其中超滤法1的回收率在3种浓度的样本中均较高,铝盐沉淀法和阴离子膜吸附—洗脱法次之,PEG沉淀法在高浓度样本中回收率较高,但在低、中浓度的样本中回收率均较低,而超滤法2的病毒回收率在3种浓度的样本中均为最低。对5种方法富集后样本中MS2的Ct值进行比较,3种浓度模拟样本中超滤法1 Ct值均为最小,中、高浓度模拟水样中5种富集方法的MS2 Ct值之间差异均无统计学意义(均P>0.05),低浓度模拟水样中仅超滤法1与超滤法2富集后的MS2 Ct值差异有统计学意义(Z=16.000,P=0.016)。结论综合考虑各方法操作简便性、操作时长及富集后病毒回收率,超滤法用于富集生活饮用水中的病毒效果最佳。ObjectiveTo compare the enrichment effects of ultrafiltration,polyethylene glycol(PEG)precipitation,aluminum salt precipitation,and anionic membrane adsorption-elution on viruses in drinking water.MethodsUsing phage MS2 as the target virus,three different concentrations of drinking water samples were prepared,and the samples were enriched by ultrafiltration 1,ultrafiltration 2,PEG precipitation,aluminum salt precipitation,and anionic membrane adsorption-elution method,respectively.Real-time fluorescence quantitative reverse transcription-polymerase chain reaction(RT-qPCR)was used to quantify MS2 nucleic acid in pre and post concentrated samples and the recovery rates of MS2 in samples with high,medium and low concentrations were compared among the five methods.ResultsComparing the MS2 enrichment recovery rates of individual enrichment method in water samples of different concentrations,ultrafiltration method 1,PEG precipitation method,aluminum salt precipitation method,and membrane adsorption-elution method were not affected by the sample concentration,and the differences of the recovery rates for the three concentration water samples among the four methods were not statistically significant(P>0.05).The MS2 enrichment recovery rates of the five enrichment methods were significantly different in all concentration samples(P<0.05).The recovery rates of ultrafiltration method 1 were higher in all three concentration samples,followed by aluminum salt precipitation and anionic membrane adsorption-elution,PEG precipitation were higher in high concentration samples,but lower in low and medium concentration samples,and the recovery rates of ultrafiltration method 2 were the lowest in all three concentration samples.Comparing the Ct values of MS2 in the enriched samples by five methods,the Ct values of ultrafiltration method 1 were the smallest in the three concentration water samples.There was no statistically significant difference in MS2 Ct values among the five enrichment methods in the medium and high concentration wa

关 键 词：病毒富集 超滤法 铝盐沉淀法 PEG沉淀法 阴离子膜吸附—洗脱法 

分 类 号：X70[环境科学与工程—环境工程]