基于数字PCR方法的污水中新冠病毒监测  

Monitoring of SARS-CoV-2 in sewage based on digital PCR method

作　　者：杨喜 程雁鹏 杜琛 彭悦景 黄定杰 韦斌财 石秀园 岳芝娇 郜晨曦 陈丽丽 李迎慧[2] 扈庆华 YANG Xi;CHENG Yan-peng;DU Chen;PENG Yue-jing;HUANG Ding-jie;WEI Bin-cai;SHI Xiu-yuan;YUE Zhi-jiao;GAO Chen-xi;CHEN Li-li;LI Ying-hui;HU Qing-hua(Department of Public Health Laboratory Sciences,School of Public Health,Hengyang Medical School,University of South China,Hengyang 421001,China;不详)

机构地区：[1]南华大学衡阳医学院,公共卫生学院,卫生检验与检疫系,湖南衡阳421001 [2]深圳市疾病预防控制中心,广东深圳518055 [3]南方医科大学 [4]山西医科大学 [5]南方科技大学

出　　处：《现代预防医学》2024年第21期3991-3995,共5页Modern Preventive Medicine

基　　金：国家自然科学基金(82373704);深圳市医学重点学科(公共卫生重点专科)(SZXK064);深圳市引进高层次医学团队项目“三名工程”(SZSM202311015);深圳市科技计划项目(KCXFZ20230731093959008)。

摘　　要：目的 探索基于数字PCR检测深圳市污水中新冠病毒浓度的方法,实现实时监测污水中新冠病毒核酸,并通过核酸浓度反映人群新冠病毒感染流行情况。方法 建立污水中新冠病毒的数字PCR检测体系并测定其最低检出限;于2023年4—7月在深圳市南山区、福田区共六个污水处理厂的进水口采集24 h混合污水样本,每周采集两次;采用改良聚乙二醇沉淀法对污水样本进行病毒富集浓缩,并使用建立的逆转录数字PCR(RT-dPCR)体系进行新冠病毒核酸定量检测。同期收集污水监测覆盖区域人群新冠病毒感染病例数据,与污水中新冠病毒核酸浓度进行相关性分析。结果研究建立了新冠病毒核酸检测的RT-dPCR方法,其最低检出限为1.00 copies/μl。在2023年4—7月期间共采集污水样本162份,新冠病毒核酸阳性检出率为96.3%。南山区新冠病毒核酸浓度范围为1.00×10~3~1.08×10~6copies/L,福田区为1.44×10~3~1.40×10~6copies/L,污水中病毒核酸浓度于5月16日达到峰值。相关性分析表明污水新冠病毒核酸浓度与人群新冠感染病例显著相关(南山区r=0.77,P<0.001;福田区r=0.80,P<0.001)。结论 数字PCR方法适用于定量检测污水中低浓度的新冠病毒核酸,可实时监测新冠疫情的流行情况,为公共卫生管理和干预提供科学依据。Objective To explore a method for monitoring the concentration of SARS-CoV-2 in sewage in Shenzhen using digital PCR,enabling real-time detection of viral nucleic acids in sewage and reflecting the epidemiological status of COVID-19 infections in the population through nucleic acid concentration.Methods A digital PCR detection system for SARS-CoV-2 in sewage was established,and its minimum detection limit was determined.From April to July 2023,24-hour composite sewage samples were collected twice weekly from the inflow of six sewage treatment plants in Nanshan and Futian districts of Shenzhen.The modified polyethylene glycol precipitation method was used for viral concentration enrichment,followed by quantitative detection of SARS-CoV-2 nucleic acids using the established reverse transcription digital PCR(RTd PCR) method.Concurrently,data on COVID-19 infection cases in the monitored sewage areas were collected for correlation analysis with the nucleic acid concentrations in the sewage.Results The RT-dPCR method for SARS-CoV-2 nucleic acid detection was successfully established,with a minimum detection limit of 1.00 copies/μl.A total of 162 sewage samples were collected from April to July 2023,with a positive detection rate of 96.3% for SARS-CoV-2 nucleic acids.The concentration of SARS-CoV-2 nucleic acids in Nanshan district ranged from 1.00 ×10~3 to 1.08 ×10~6 copies/L,while in Futian district it ranged from 1.44×10~3 to 1.40×10~6 copies/L,with peak concentrations observed on May 16.Correlation analysis indicated a significant association between sewage SARS-CoV-2 nucleic acid concentrations and the number of COVID-19 infection cases in the population(Nanshan district r=0.77,P<0.001;Futian district r=0.80,P<0.001).Conclusion The digital PCR method is suitable for the quantitative detection of low concentrations of SARS-CoV-2 nucleic acids in sewage,allowing for real-time monitoring of the COVID-19 epidemic,thus providing a scientific basis for public health management and intervention.

关 键 词：污水监测 新型冠状病毒 数字PCR 新型冠状病毒感染 

分 类 号：R211[医药卫生—中医学] R123.1