2022—2023年深圳污水中诺如病毒的监测分析  

A wastewater-based epidemiology monitoring study for norovirus in Shenzhen,2022-2023

作　　者：潘荣玲 旷翠萍[1] 陈仕菊 张海龙[3] 万佳 王礼法[1] 黄志刚 PAN Rong-ling;KUANG Cui-ping;CHEN Shi-ju;ZHANG Hai-long;WAN Jia;WANG Li-fa;HUANG Zhi-gang(Luohu District Center for Disease Control and Prevention,Shenzhen,Guangdong 518020,China;不详)

机构地区：[1]深圳市罗湖区疾病预防控制中心,广东深圳518020 [2]广东医科大学公共卫生学院,广东东莞523808 [3]深圳市疾病预防控制中心

出　　处：《现代预防医学》2024年第13期2348-2353,共6页Modern Preventive Medicine

基　　金：深圳市科技计划(JCYJ20210324130215038)。

摘　　要：目的 为了探索诺如病毒的分布和时间动态变化特征,初步量化污水中诺如病毒基因I型和II型 (GI和GII) RNA浓度,以构建污水中诺如病毒定量检测数据库,为进一步开展基于污水流行病学诺如病毒监测预测人群感染的研究提供基本资料与数据支撑。方法 在深圳市设立5个污水采样点,进行为期一年的定性定量监测。从2022年8月—2023年8月,每周规律采样。 采用逆转录PCR( reverse transcription-PCR , RT-PCR)方法,对诺如病毒(GI和GII)进行检测和定性分析;采用圆盘电泳过滤器和聚乙二醇沉淀法,提取核酸;采用定量逆转录 -聚合酶链反应 (quantitative reverse transcription-polymerase chain reaction,RT-qPCR)进行定量分析。诺如病毒 RNA载量进行log10转化,数值归一化处理;率的比较采用χ2检验或Fisher确切概率法;对定量数据(不符合正态分布)比较采用Wilcoxon符号秩和检验。结果 在5个污水处理厂采集的260份污水中,诺如病毒 GI型、GII型的总检出率分别为88.85%、91.15%,其中,96.5%样品 同时检测出GI、GII型。研究期间,诺如病毒在2月和3月达到高峰,GI、GII型最高值达到108.67 GC/L和108.06 GC/L。诺如病毒在原始废水样品中广泛分布,GI、GII型分别为83.3%~100%和91.6%~100%,其中,诺如病毒GI、GII型RNA载量范围分别为3.8~6.66 log10 GC/L、3.8~7.3 log10 GC/L,均值为106.11 GC/L、106.15 GC/L。结论 通过污水中诺如病毒的监测,证实诺如病毒常年存在,全年四季的病毒浓度差异及其趋势特征符合诺如病毒季节性流行病学特征。探索利用污水流行病学方法来提示诺如病毒在人群中的流行病学特征,是对目前主要基于确诊病例被动监测的重要补充。Objective To investigate the distribution and temporal dynamics of Norovirus(NoV)in wastewater through preliminary quantification of genotypes I and II(GI and GII)RNA concentrations.The goal was to establish a database for quantitative detection of NoV in wastewater and provide foundational data for further research on predicting population infections using wastewater-based epidemiological surveillance of Norovirus.Methods Five sewage sampling sites were established in Shenzhen for qualitative and quantitative surveillance over one year.Weekly samples were collected from September 2022 to August 2023,totaling 260 samples over 52 weeks.Real-time fluorescence reverse transcription-polymerase chain reaction(Real-time RT-PCR)was employed to detect and analyze NoV(including GI and GII).Sample concentration involved two steps:disc electrophoresis filtration and polyethylene glycol precipitation,followed by nucleic acid extraction and quantitative polymerase chain reaction analysis.Results The overall detection rates of NoV GI and NoV GII were 84.62%and 89.5%,respectively,with 96.5%detecting both genotypes simultaneously.NoV concentrations in wastewater ranged from 103.56 to 108.67 genomic copies per liter(GC/L)for NoV GI and from 104.63 to 108.06 GC/L for NoV GII.The highest RNA concentration detected was 108.67 GC/L.NoV was widely distributed in original wastewater samples(83.3%NoV GI and 91.6%NoV GII),with viral loads of 3.86 log10 GC/L(GI)and 3.87 log10 GC/L(GII),averaging 6.11 log10 GC/L(GI)and 6.15 log10 GC/L(GII).Conclusion Continuous monitoring of Norovirus in sewage confirmed its presence throughout the year,with concentrations varying seasonally in accordance with epidemiological trends.Utilizing wastewater-based epidemiology offers insights into the epidemiological characteristics of Norovirus,complementing existing surveillance methods reliant on confirmed cases.

关 键 词：污水 诺如病毒 定量逆转录聚合酶链反应(q-PCR) 多重实时荧光(RT-PCR) 病毒载量 

分 类 号：X508[环境科学与工程—环境工程] R114[医药卫生—卫生毒理学]