Article(id=1316714455040377486, tenantId=1146029695717560320, journalId=1192105938417971205, issueId=1316714454184739470, articleNumber=null, orderNo=null, doi=10.13343/j.cnki.wsxb.20260172, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=research-article, receivedDate=1772467200000, receivedDateStr=2026-03-03, revisedDate=null, revisedDateStr=null, acceptedDate=1774454400000, acceptedDateStr=2026-03-26, onlineDate=1791858789452, onlineDateStr=2026-10-13, pubDate=1791043200000, pubDateStr=2026-10-04, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1791858789452, onlineIssueDateStr=2026-10-13, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1791858789452, creator=13701087609, updateTime=1791858789452, updator=13701087609, issue=Issue{id=1316714454184739470, tenantId=1146029695717560320, journalId=1192105938417971205, year='2026', volume='66', issue='10', pageStart='4661', pageEnd='5310', issueExtLink='null', onlineDate='null', pubDate='1791043200000', pubDateStr='2026-10-04', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=1, specialIssue=null, createTime=1791858789249, creator='13701087609', updateTime=1791885400527, updator='13701087609', preIssue=null, nextIssue=null, articleTotal=null, ext={EN=IssueExt(id=1316826070066774161, tenantId=1146029695717560320, journalId=1192105938417971205, issueId=1316714454184739470, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1316826070066774162, tenantId=1146029695717560320, journalId=1192105938417971205, issueId=1316714454184739470, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null, downloadFileDto=null}, startPage=4679, endPage=4693, ext={EN=ArticleExt(id=1316714455313007247, articleId=1316714455040377486, tenantId=1146029695717560320, journalId=1192105938417971205, language=EN, title=Research progress in quantitative monitoring and risk assessment of pathogenic microorganisms in drinking water, columnId=1192149543727808575, journalTitle=Acta Microbiologica Sinica, columnName=Review, runingTitle=null, highlight=null, articleAbstract=

The health risks posed by pathogenic microbial contamination in drinking water are becoming increasingly apparent. Following exposure to pathogens via water, the human body may experience reactions ranging from infection and illness to death. However, the current Standards for Drinking Water Quality (GB 5749—2022) primarily rely on indicator microorganisms to evaluate water quality, lacking precise targets for pathogen control and load levels, which makes it difficult to accurately assess the actual hazards posed by microorganisms in drinking water. Therefore, clarifying the relationship between the intake dose of pathogenic microorganisms in drinking water and the risk of human infection is of great significance for safeguarding public health. Quantitative microbial risk assessment (QMRA) can be employed to evaluate the health risks posed by pathogenic microorganisms in drinking water. It is currently combined with high-throughput molecular technologies such as real-time quantitative PCR, digital quantitative PCR, metagenomics, and metaviromics, overcoming the limitations of conventional culture-based methods in identifying pathogenic hazards and demonstrating significant advantages in pathogen hazard identification and multidimensional traceability. This paper reviews the research progress in the quantitative monitoring and risk assessment of pathogenic microorganisms in drinking water. On the basis of the distribution patterns and quantitative detection of pathogenic microorganisms, QMRA is employed to conduct risk prediction, screen high-risk pathogens, and identify critical control points, and thus a systematic approach for precise prevention and control and full-chain governance is ultimately established. This approach integrates multi-omics technologies, big data analysis, and health impact assessment, achieving a transition in water quality safety management from reactive response to proactive prevention and providing robust scientific support for ensuring drinking water safety and the health of vulnerable populations.

, authors=Yunchu SHEN1, 2, Ling CHEN2, Qingping WU2, Qihui GU2, *, Xinyu ZHAO3, *, authorsList=Yunchu SHEN, Ling CHEN, Qingping WU, Qihui GU, Xinyu ZHAO, authorCompany=null, correspAuthors=Qihui GU, Xinyu ZHAO, authorNote=null, correspAuthorsNote=
*E-mail: ZHAO Xinyu,
GU Qihui,
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饮用水病原微生物污染带来的健康风险日益凸显。病原微生物经水途径暴露后,人体的反应可能有感染、致病、死亡等。然而,现行《生活饮用水卫生标准》(GB 5749—2022)主要依赖指示菌评价水质,缺乏精准的病原微生物防控对象和负载量,难以准确评估饮用水中微生物的实际危害。因此,明确饮用水中病原微生物的摄入剂量与人体感染风险之间的关系对保障公共卫生安全具有重要意义。定量微生物风险评估(quantitative microbial risk assessment, QMRA)可用于评估饮用水中病原微生物的健康风险,目前已与实时荧光定量PCR、数字荧光定量PCR、宏基因组和宏病毒组等高通量分子技术联用,突破了传统培养法识别病原微生物危害的局限,在病原微生物危害识别与多维溯源中展现出显著优势。本文综述了饮用水中病原微生物的定量监测和风险评估研究进展,以病原微生物的赋存规律与定量检测为基础,结合QMRA进一步开展风险预测、高风险病原筛查与关键控制点识别,最终建立起精准防控与全链条治理的系统方案。该方案交叉融合多组学技术、大数据分析与健康效应评估,实现了水质安全管理由“被动应对”向“主动预防”的跨越,为保障饮用水安全及易感人群健康提供了有力的科学支撑。

, authors=沈沄楚1, 2, 陈玲2, 吴清平2, 古其会2, *, 赵昕宇3, *, authorsList=沈沄楚, 陈玲, 吴清平, 古其会, 赵昕宇, authorCompany=null, correspAuthors=古其会, 赵昕宇, authorNote=

作者贡献声明

沈沄楚:论文构思、论文撰写;陈玲:文献调研、数据分析;吴清平:问题提出、论文修改;古其会:论文构思、论文修改;赵昕宇:问题提出、参与讨论。

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Discussion on the application of improved method for water quality concentration prediction in Taizi River[J]. 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Microbiological monitoring techniques for drinking water

, figureFileSmall=null, figureFileBig=null, tableContent=
MethodTesting featuresReferences
Traditional cultivation methodsStandardized for regulatory and routine monitoring, low-cost and easy to use, but prone to false negatives, with long testing cycles and results highly susceptible to processing variables[27-28,30]
dPCR and viability-qPCRdPCR allows absolute quantification with strong anti-interference, suitable for low-concentration samples. Viability-qPCR and ci-qPCR distinguish live and dead cells and viral activity, indicating infectivity. However, equipment is costly, conditions need optimization, and differentiation may be incomplete[5-6]
High-throughput sequencing and metagenomicsComprehensive analysis of microbial communities, functional genes and pathogens. Very high cost; hard to link activity with infection risk; difficult to meet regulatory standards[29,33,35-36,49]
Online flow cytometry and ATP monitoringOnline, rapid monitoring of total and intact cells and biological activity. Thresholds and classification are subjective; alignment with regulations unclear[7,38-39]
Portable and novel biosensorsFast, portable, suitable for on-site use; aptamer sensors allow sensitive quantification of target bacteria. Lacks on-site reference materials and quality control; mostly at prototype stage with few standardized applications[8,42-45,50]
), ArticleFig(id=1316829713738355591, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714455040377486, language=CN, label=表1, caption=

饮用水微生物监测技术

, figureFileSmall=null, figureFileBig=null, tableContent=
MethodTesting featuresReferences
Traditional cultivation methodsStandardized for regulatory and routine monitoring, low-cost and easy to use, but prone to false negatives, with long testing cycles and results highly susceptible to processing variables[27-28,30]
dPCR and viability-qPCRdPCR allows absolute quantification with strong anti-interference, suitable for low-concentration samples. Viability-qPCR and ci-qPCR distinguish live and dead cells and viral activity, indicating infectivity. However, equipment is costly, conditions need optimization, and differentiation may be incomplete[5-6]
High-throughput sequencing and metagenomicsComprehensive analysis of microbial communities, functional genes and pathogens. Very high cost; hard to link activity with infection risk; difficult to meet regulatory standards[29,33,35-36,49]
Online flow cytometry and ATP monitoringOnline, rapid monitoring of total and intact cells and biological activity. Thresholds and classification are subjective; alignment with regulations unclear[7,38-39]
Portable and novel biosensorsFast, portable, suitable for on-site use; aptamer sensors allow sensitive quantification of target bacteria. Lacks on-site reference materials and quality control; mostly at prototype stage with few standardized applications[8,42-45,50]
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饮用水中病原微生物的定量监测和风险评估研究进展
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沈沄楚 1, 2 , 陈玲 2 , 吴清平 2 , 古其会 2, * , 赵昕宇 3, *
微生物学报 | 综述 2026,66(10): 4679-4693
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微生物学报 |综述 2026 , 66 (10) : 4679 -4693
饮用水中病原微生物的定量监测和风险评估研究进展
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沈沄楚1, 2, 陈玲2, 吴清平2, 古其会2, * , 赵昕宇3, *
作者信息
  • 1.华南农业大学 食品学院,广东 广州
  • 2.广东省科学院微生物研究所,华南应用微生物国家重点实验室,广东省微生物安全与健康重点实验室,国家卫健委微生物食品营养与安全科技创新平台,国家市场监督管理总局重点实验室(食品微生物安全 大数据技术),广东 广州
  • 3.广东环凯信息科技有限公司,广东 广州
通讯作者:
古其会, 赵昕宇
作者简介:

作者贡献声明

沈沄楚:论文构思、论文撰写;陈玲:文献调研、数据分析;吴清平:问题提出、论文修改;古其会:论文构思、论文修改;赵昕宇:问题提出、参与讨论。

Research progress in quantitative monitoring and risk assessment of pathogenic microorganisms in drinking water
Yunchu SHEN1, 2, Ling CHEN2, Qingping WU2, Qihui GU2, * , Xinyu ZHAO3, *
Affiliations
  • 1.College of Food Science, South China Agricultural University, Guangzhou, Guangdong, China
  • 2.Institute of Microbiology, Guangdong Academy of Sciences, State Key Laboratory of Applied Microbiology Southern China, Guangdong Provincial Key Laboratory of Microbial Safety and Health, National Health Commission Science and Technology Innovation Platform for Nutrition and Safety of Microbial Food, Key Laboratory of Big Data Technologies for Food Microbiological Safety, State Administration for Market Regulation, Guangzhou, Guangdong, China
  • 3.Guangdong Huankai Biotechnology Co. , Ltd. , Guangzhou, Guangdong, China
  • Corresponding Author:
    *E-mail: ZHAO Xinyu,
    GU Qihui,
出版时间: 2026-10-04 doi: 10.13343/j.cnki.wsxb.20260172
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饮用水病原微生物污染带来的健康风险日益凸显。病原微生物经水途径暴露后,人体的反应可能有感染、致病、死亡等。然而,现行《生活饮用水卫生标准》(GB 5749—2022)主要依赖指示菌评价水质,缺乏精准的病原微生物防控对象和负载量,难以准确评估饮用水中微生物的实际危害。因此,明确饮用水中病原微生物的摄入剂量与人体感染风险之间的关系对保障公共卫生安全具有重要意义。定量微生物风险评估(quantitative microbial risk assessment, QMRA)可用于评估饮用水中病原微生物的健康风险,目前已与实时荧光定量PCR、数字荧光定量PCR、宏基因组和宏病毒组等高通量分子技术联用,突破了传统培养法识别病原微生物危害的局限,在病原微生物危害识别与多维溯源中展现出显著优势。本文综述了饮用水中病原微生物的定量监测和风险评估研究进展,以病原微生物的赋存规律与定量检测为基础,结合QMRA进一步开展风险预测、高风险病原筛查与关键控制点识别,最终建立起精准防控与全链条治理的系统方案。该方案交叉融合多组学技术、大数据分析与健康效应评估,实现了水质安全管理由“被动应对”向“主动预防”的跨越,为保障饮用水安全及易感人群健康提供了有力的科学支撑。

饮用水  /  病原微生物  /  微生物定量检测  /  定量微生物风险评估(QMRA)

The health risks posed by pathogenic microbial contamination in drinking water are becoming increasingly apparent. Following exposure to pathogens via water, the human body may experience reactions ranging from infection and illness to death. However, the current Standards for Drinking Water Quality (GB 5749—2022) primarily rely on indicator microorganisms to evaluate water quality, lacking precise targets for pathogen control and load levels, which makes it difficult to accurately assess the actual hazards posed by microorganisms in drinking water. Therefore, clarifying the relationship between the intake dose of pathogenic microorganisms in drinking water and the risk of human infection is of great significance for safeguarding public health. Quantitative microbial risk assessment (QMRA) can be employed to evaluate the health risks posed by pathogenic microorganisms in drinking water. It is currently combined with high-throughput molecular technologies such as real-time quantitative PCR, digital quantitative PCR, metagenomics, and metaviromics, overcoming the limitations of conventional culture-based methods in identifying pathogenic hazards and demonstrating significant advantages in pathogen hazard identification and multidimensional traceability. This paper reviews the research progress in the quantitative monitoring and risk assessment of pathogenic microorganisms in drinking water. On the basis of the distribution patterns and quantitative detection of pathogenic microorganisms, QMRA is employed to conduct risk prediction, screen high-risk pathogens, and identify critical control points, and thus a systematic approach for precise prevention and control and full-chain governance is ultimately established. This approach integrates multi-omics technologies, big data analysis, and health impact assessment, achieving a transition in water quality safety management from reactive response to proactive prevention and providing robust scientific support for ensuring drinking water safety and the health of vulnerable populations.

drinking water  /  pathogenic microorganisms  /  quantitative microbial detection  /  quantitative microbial risk assessment (QMRA)
沈沄楚, 陈玲, 吴清平, 古其会, 赵昕宇. 饮用水中病原微生物的定量监测和风险评估研究进展. 微生物学报, 2026 , 66 (10) : 4679 -4693 . DOI: 10.13343/j.cnki.wsxb.20260172
Yunchu SHEN, Ling CHEN, Qingping WU, Qihui GU, Xinyu ZHAO. Research progress in quantitative monitoring and risk assessment of pathogenic microorganisms in drinking water[J]. Acta Microbiologica Sinica, 2026 , 66 (10) : 4679 -4693 . DOI: 10.13343/j.cnki.wsxb.20260172
水是人类赖以生存和发展的根本,饮用水安全直接关乎人们的身体健康。近些年,介水感染事件频发,病原微生物污染的水源是人类致病和致死的主要风险因素之一[1]。饮用水中的病原微生物污染已然成为公共卫生安全的重大挑战。嗜肺军团菌(Legionella pneumophila)、铜绿假单胞菌(Pseudomonas aeruginosa)与非结核分枝杆菌(non-tuberculous Mycobacteria, NTM)等机会致病菌(opportunistic pathogens, OPs)在配水系统与建筑末梢生物膜中被频繁检出[2],隐孢子虫(Cryptosporidium)和诺如病毒(human Norovirus, HuNoV)在极端降雨、管网负压入渗或处理波动时也可造成公共危害事件[3]。因此,了解这些病原微生物的污染水平、分布特征和接触风险是预防水传播疾病的关键条件。现行监测体系仍以传统检测技术和基于分子生物学的高新检测技术为主[4-9]。尽管这些方法在合规性评价中起着重要作用,但它们仍存在分析时间较长,对低浓度或休眠态微生物不敏感,难以直接反映微生物活性,缺乏实施标准规范,以及与健康风险的定量关系不明确等局限。定量微生物风险评估(quantitative microbial risk assessment, QMRA)在风险管理决策中正日益受到推崇,近年来的关键进展集中在表征环节的可解释性与可决策性[10]。在此基础上,现有监测技术应从“单一指标合规”转向“面向健康风险的组合监测”。培养法可作为法规底线与长期趋势判别的基准[11],应按目的与场景联合培养法、分子法与在线监测等数据,将工艺变量与检测结果联动[12],将检测数据转化为可用于QMRA的暴露参数与风险量化输出[13]。
面对我国饮用水微生物风险检测和防控能力需要提高,检测方法多、杂,普遍存在缺陷,无法面对复杂的实际情况,定量微生物风险评估难以落实、难以本土化的情况,将传统微生物检测结果与基于分子生物学的高新检测结果实现等效转换并与QMRA相结合,建立本土化评估指标,贯通“阈值确定-干预实施-量化复评”的闭环健康风险监测逻辑,构建可复评可追溯的饮用水生物风险治理框架,对于有效监管饮用水中的微生物污染至关重要,是确保饮用水安全、实现病原微生物管控的重要手段。
我国现行国家标准《生活饮用水卫生标准》(GB 5749—2022)对微生物安全提出了明确硬约束:不得检出总大肠菌群与大肠埃希氏菌(Escherichia coli),并限制菌落总数,以保障供水末端的卫生安全边界[3]。世界卫生组织(World Health Organization, WHO)《饮用水水质指南(第三版及其增编)》指出,饮用水微生物风险控制应以健康为基础,通过建立健康目标并实施水安全计划(water safety plan, WSP)进行持续管理,核心目标是确保饮用水中不含致病微生物;在管理措施上,建议同步开展常规指标监测和系统性风险管理[1]。
过去5年,国家与地方监测资料显示饮用水微生物指标的总体合规度在提升,但在农村集中供水、小型水厂、末梢水以及季节等维度仍存在明显差异。以重庆市巴南区为例,2020-2022年共监测460份样品,微生物指标在各类指标中合格率最低(83.26%);不同水源类型与消毒方式的达标率差异显著:丰水期江河水源(96.88%)高于水库(85.06%)和其他水源(54.17%);枯水期采用液氯或次氯酸钠的达标率高于二氧化氯(χ2=14.03, P<0.001);同一水源在枯水期采用复合二氧化氯的达标率(90.63%),高于丰水期(81.25%)[14]。这些结果提示,水源属性、季节变化与消毒运行是影响微生物达标的关键变量,需要在监测设计与运维管理中加以分层与针对性控制。与此同时,来自我国农村集中供水的一年期调查表明:即便在集中供水体系中,机会致病菌相关基因仍普遍检出,且在夏秋季受残氯不足与浊度升高影响而出现上升;在小型集中模式下,一些病原微生物如假单胞菌属(Pseudomonas)、鸟分枝杆菌(Mycobacterium avium)、棘阿米巴属(Acanthamoeba)与指示菌如大肠埃希氏菌的基因拷贝数在个别时段可达103-107级别,提示季节、管网稳定性和水质理化参数共同作用的微生物变化的重要性,建议在时间、空间与水质理化参数等维度进行更细化的监测与干预[15]。
在集中供水与管网系统中,传统粪便指示菌、配水系统病原微生物以及典型水源性肠道病原微生物共同构成了饮用水微生物风险谱系。总大肠菌群与大肠埃希氏菌长期被用作粪便污染和饮用水配水管网系统微生物污染的主要指示菌。总大肠菌群检出反映水体受微生物污染的整体状况,预示肠道病原微生物的潜在存在风险;大肠埃希氏菌为粪便污染的特异性指示菌,其检出直接表明水体受人畜粪便污染,易引发急性肠道感染等健康危害,二者的存在均会增加饮用水病原微生物暴露导致的人群患病风险,本文总结了水源性病原微生物、暴露途径及国内外相关饮用水标准概览(图1)。我国《生活饮用水卫生标准》(GB 5749—2022)要求总大肠菌群和大肠埃希氏菌不得检出,并规定菌落总数限值[3],但在农村集中供水与部分初级处理工艺水厂中微生物去除效果有限,这2种病原微生物均具有较高的检出风险,威胁人们的身体健康。
与粪便指示菌不同,嗜肺军团菌、铜绿假单胞菌和非结核分枝杆菌等机会致病菌多在配水管网和建筑末梢水的生物膜中长期定植,典型暴露途径是淋浴、冷却塔或末梢用水形成的含菌气溶胶,而非单纯饮水。嗜肺军团菌为耐低营养的革兰阴性杆菌,可通过吸入含菌气溶胶引起军团病和庞蒂亚克热,摄入被污染饮水通常并非主要暴露途径。欧盟《饮用水指令》将军团菌列为监测和管理参数,用户龙头常以约1 000 CFU/L作为干预阈值[16]。铜绿假单胞菌为革兰阴性好氧杆菌,对消毒具有较强耐受能力,易在配水系统与建筑末梢生物膜中持续存在[17],是创伤、重症和免疫低下人群的重要机会致病菌[18],可经接触受污染水或气溶胶进入伤口、呼吸道、眼和耳等部位[17]。研究表明,供水管网中的铜绿假单胞菌可以以活的非可培养状态(viable but non-culturable, VBNC)持续存在[18],而传统培养方法通常低估其真实负荷[19];多国医院饮用水及末梢系统中分离出的对氯耐受铜绿假单胞菌也提示单一的化学消毒效果有限[17]。非结核分枝杆菌具有细胞壁疏水、耐消毒及耐营养贫乏等特性,在配水系统和建筑末梢生物膜中尤为顽固,主要通过含菌气溶胶误吸暴露,免疫低下与基础肺病人群风险更高。系统研究显示,低余氯、温热和长水龄均可促进非结核分枝杆菌丰度提升,建筑末梢滞留点通常浓度最高[3,20];院内和长期护理设施中也有与供水系统相关的非结核分枝杆菌获得性感染报道。
隐孢子虫和诺如病毒则是典型的水源性肠道病原微生物。隐孢子虫卵囊耐氯、感染剂量低,可通过摄入受污染饮用水引起急性胃肠炎[21]。暴雨水源污染、管网破损、负压入渗或处理工艺故障时,其暴露风险显著升高。WHO相关文件和2022版《饮用水水质指南》提出健康阈值和对原虫的对数去除(log reduction value, LRV)或灭活绩效控制隐孢子虫风险,强调源水防护和多屏障工艺[22]。实验室检测方面,目前多采用大体积水样浓缩后进行免疫磁分离(immunomagnetic separation, IMS)和荧光显微镜计数,或利用实时荧光定量PCR (quantitative real-time PCR, qPCR)进行物种和基因型鉴定。
诺如病毒具极低感染剂量、环境稳定性强和多基因型的特征[23],是全球急性胃肠炎的首要病原之一,在源水受粪便污染、管网破损或消毒失效时可进入饮用水系统,通过直接饮水或食物传播引发暴发。2022年意大利北部公共供水及2024年印度多地的调查均证实诺如病毒污染饮用水与区域暴发密切相关[24-25]。美国疾病控制与预防中心(Centers for Disease Control and Prevention, CDC)国家食源性疾病暴发监测系统(National Outbreak Reporting System, NORS)数据显示,诺如病毒暴发处于高位运行,并提示饮水相关暴发通常被误归为食源性而被漏报,需要加强水样核酸检测与暴发溯源。我国疾病控制中心的指导原则是一旦从水中检出诺如病毒核酸阳性,且与患者感染毒株基因型高度同源,即判定为水型暴发,必须立即切断供水并进行终末消毒。本团队研究人员在食品安全国家标准的规划、组织及制修订工作中,首次推动将诺如病毒检测方法纳入国家标准[26]。
饮用水环境中存在着多种病原微生物,包括粪便指示菌、机会性致病菌、原虫和病毒等,可能引起水媒传染病的发生和流行,可导致呕吐、腹泻、腹痛等胃肠道症状,也可引起发热、肌肉酸痛、关节痛等感染症状,甚至可造成感染者休克或者死亡,直接威胁到公共卫生健康和生态稳定。掌握饮用水中病原微生物的污染水平、分布特征及暴露风险,是防控介水传染病、保障饮用水安全的关键前提,因此对这些病原微生物的评估检测至关重要。从大肠埃希氏菌和总大肠菌群等传统指示菌到军团菌、铜绿假单胞菌、非结核分枝杆菌等机会致病菌,再到隐孢子虫和诺如病毒等典型水源性病原微生物,目前现行监测体系主要以传统培养法、分子生物学检测方法和高通量测序方法为主。上述方法虽然在法规合规性评价中发挥了重要作用,但普遍存在检测耗时长、对低水平或短时污染以及微生物休眠状态不敏感、难以直接反映微生物活性和复杂的微生物群落组成并与健康风险定量关联不清等局限。这些不足在一定程度上限制了对处理工艺波动、管网二次污染和末梢水系统失稳的实时识别,也为发展更快速、高灵敏、可在线或原位应用的新型饮用水微生物检测技术提供了现实需求和研究方向。
以膜过滤(membrane filtration, MF)、多管发酵(multiple-tube fermentation technique, MTFT)与异养菌落总数(heterotrophic plate count, HPC)为代表的培养法是微生物检测传统技术。近年综述与方法学评估指出:针对总大肠菌群和耐热大肠菌群、大肠埃希氏菌的膜过滤与发酵法在低成本、标准化方面具有优势[27-28],但对受损细胞、VBNC[29]与低浓度末梢样品存在假阴性和灵敏度受限等问题,需要通过前增菌、选择性复苏和优化采样体积与介质来改进。传统培养法研究提示,采样方式(滤膜或液体直取)、水龄、残氯、季节、水源等工艺变量常与培养结果的离散性结合分析,使得检测方法的表现与具体采样点的环境特征紧密关联[30]。
数字荧光定量PCR (digital PCR, dPCR)的绝对定量和抗抑制优势,已在健康相关水微生物中得到系统评估,常用以弥补qPCR在低浓度、抑制物存在下的偏差[6];本团队通过逆转录实时PCR (real-time reverse transcription quantitative PCR, RT-qPCR)比较实体市场与电商平台所售太平洋牡蛎(Crassostrea gigas)中HuNoV和人源札幌病毒 (human sapovirus, HuSaV)的污染水平,体现牡蛎中食源性病毒的高污染率[31];存活态实时荧光定量聚合酶链式反应(viability quantitative real‑time polymerase chain reaction, viability-qPCR)或衣壳完整性实时荧光定量聚合酶链式反应(capsid integrity quantitative real‑time polymerase chain reaction, ci-qPCR)用于排除死细胞和失活衣壳信号,从而使分子结果更接近“潜在感染性”。本团队针对环境中的铜绿假单胞菌开发了一款多重PCR检测技术,即使非目标分析物细胞浓度达到109 CFU/mL,也具有很强的抗干扰能力[32]。最新元分析结果提示,在嗜肺军团菌的qPCR的检测结果应用于定量微生物风险评估前,采用qPCR培养1:1的保守换算系数[7],并在消毒显著的体系考虑viability-qPCR以降低高估风险。综合来看,dPCR和viability-qPCR结合传统培养的互证框架最有利于风险评估输入与应急处置决策。
宏基因组和全长16S rRNA基因测序可用于解析配水系统核心群落、事件扰动以及潜在病原相关特征;基因或基因组中心分析在多座水厂应用中已用于推断功能与传播。2024-2025年多项研究将耐药基因组与宿主或可移动遗传元件(mobile genetic elements, MGEs)一并解析[33];本团队对包装饮用水处理系统的宏基因组拼装展示了处理单元中的耐药基因(antibiotic resistance genes, ARGs)和耐药细菌(antibiotic-resistant bacteria, ARB)的迁移,是首个利用宏基因组学监测包装饮用水处理系统中抗生素耐药性存在及分布的研究[29],还通过宏基因组学全面揭示了饮用水处理厂中ARGs和ARB分布特征,并发现ARGs优势宿主为葡萄球菌(Staphylococcus spp.),其携带多药物ARGs最为常见[4];使用宏病毒检测技术分析地下水病毒污染情况,检测出地下水中已知病毒的微生物门类数量翻倍[34]。长读长测序(oxford nanopore technology, ONT)在耐药基因、可移动元件的关联解析上具有优势[35],已应用于追踪处理后排放进入水体的耐药基因渗透情况,以及监测配水生物膜的结构和功能特征[36]。此外,国内研究团队还对定量宏基因组进行了研究和产品开发,通过设计并引入多种外源性及合成内标(涵盖3个区段,共79条内标),有效规避内标序列与样本来源重叠导致的检测模糊,实现了高通量基因定量的精准化[37]。尽管如此,宏基因组和宏病毒等测序技术仍存在成本高、难以直接关联活性与感染风险、与法规指标衔接困难等问题,无法广泛应用。
在线流式细胞术(online flow cytometry, online-FCM)与在线三磷酸腺苷(adenosine triphosphate, ATP)监测能以分钟至小时级分辨率跟踪总细胞、完整细胞与生物稳定性,用于事件预警、消毒工艺(如臭氧)在线验证与末梢水“异常变化”判别[7]。多项研究与综述显示,与传统的异养菌落总数相比,online-FCM能够更灵敏地捕捉水处理过程中的短期水质波动;通过结合流式细胞指纹图谱或相似性指数分析(Bray-Curtis index),并参考高核酸与低核酸含量细菌的比例,可将工艺调整或污染事件与日常水质波动有效区分开来[38];同时,ATP检测结果与异养菌落总数的数据间相关性及其差异,为量化余氯与管网水生物稳定性之间的关系提供了依据[39]。然而,该技术目前仍面临若干挑战,包括门限设定与细胞分群存在主观性、算法尚未统一、不同设备间的结果可比性不足,以及与现有水质法规指标的衔接问题。
成簇规律间隔短回文重复序列-关联蛋白(clustered regularly interspaced short palindromic repeats-associated, CRISPR/Cas)结合重组酶聚合酶扩增技术(recombinase polymerase amplification, RPA)、环介导等温扩增技术(loop-mediated isothermal amplification, LAMP)[40]等恒温扩增可实现快速、特异、可现场的核酸检测;本团队有研究者将解旋酶依赖性扩增(helicase-dependent amplification, HDA)检测系统与优化的CRISPR/Cas12a整合,开发了新型“HDA-Cas12aFQ”平台,可在2 h内检测水中的铜绿假单胞菌[41];系统综述归纳了重组酶聚合酶扩增技术-成簇规律间隔短回文重复序列和关联蛋白12a (RPA-Cas12a)在病原现场检测的进展与优化思路[42]。恒温扩增也用于水与废水中短时程的批量筛查[43]。另一方面,表面增强拉曼光谱(surface-enhanced Raman scattering, SERS)便携设备配合双层过滤浓缩可在30 min内对多种水中细菌实现同步分析,检测限可达10 CFU/mL[44];适配体电化学传感[8] (如针对铜绿假单胞菌的F23适配体)在一次性电极上实现快速灵敏定量[45]。上述技术在饮水场景的现实挑战主要是基质抑制、标准物质与质量控制、设备互操作与多重化,但在应急、末端现场、物资受限条件下具有显著潜力。
微生物检测方法在饮用水中的病原微生物风险评估中发挥了重要作用,通过检测饮用水微生物量,识别微生物种类和相对丰度,能够初步评价饮用水样品的健康风险。然而,现行的检测技术普遍存在检测耗时长、对低水平或短时污染以及微生物休眠状态不敏感、难以直接反映微生物活性、与健康风险定量关联不清等局限,限制了处理工艺调整、供水管网和末梢水系统二次污染时的风险识别,这为构建新型全面的饮用水微生物检测技术提供了现实需求和研究方向。
培养法更适合作为法规底线与趋势判别工具;当用于定量风险建模(如QMRA)或复杂场景(大型配水系统末梢、生物膜与部件监测)时,需根据监测目的与场景联用培养法、分子法和在线法[46],并通过实验室间比对与转换系数来减小方法的不确定性。例如军团菌在欧盟《饮用水指令》下允许以风险控制为基础的验证思路,传统培养、快速培养、分子法可互补,2025年多实验室研究也验证了跨法一致性与差异需要统一质控与方法间转换[47]。在建筑用水场景,定量微生物风险评估正尝试将水龄、温度、残氯等工艺变量与分子、培养结果联动[46],提升暴露参数的现实性。此外,采样代表性与工艺变量记录要与数据一并存档;实验室间比对材料、质控材料与阳性、阴性对照要覆盖培养与分子;报告格式需区分qPCR基因拷贝,viability-qPCR,培养菌落形成单位(colony-forming unit, CFU)和多管发酵,并给出方法检出定量限与抑制评估;在线法需标注算法与阈值、与事件处置标准作业程序(standard operating procedure, SOP)联动;面向定量微生物风险评估的数据需给出方法不确定性与转换系数[46,48]。本文整理了饮用水微生物监测技术,并比较优劣,如表1所示。饮用水供水管网系统微生物风险受多重因素调控且时空异质性显著,单一的微生物指标管控难以满足当今社会的饮用水安全需求,构建综合性的定量微生物风险评估体系,可量化人群感染风险、支撑闭环管理,为全链条微生物安全管控提供科学依据。
水中的微生物种类繁多,介水病原微生物会对人体健康造成潜在威胁。在实际应用中通常要根据病原微生物浓度水平、感染力和致病力等因素筛选出优先考虑的目标微生物。目前存在很多有针对性的评价技术,美国国家科学院根据微生物风险评价(microbial risk assessment, MRA)建立的四步法模型(包括危害识别、暴露评估、剂量反应关系、风险表征)被世界各国或者国际组织广泛采纳。QMRA框架在此基础上开发完善,其步骤主要为危害识别、暴露评价、剂量-反应评价以及风险表征。尽管QMRA在国内食品安全研究中得到广泛应用和实施,但在饮用水病原微生物领域中仍存在因检测方式不同造成的结果差异,因此欧盟《饮用水指令》把包括原水、水厂、配水、用户端的全链条管理的风险监控写进法规[16],美国环境保护署(U.S. Environmental Protection Agency, USEPA) 2025风险基准框架给出可直接用于设定目标的参考病原、剂量反应、对数削减目标与风险对标方法[51]。以下将以QMRA的四步法为主线,阐述水环境中定量微生物风险评估技术的主要框架与相关检测技术的研究进展。
危害识别是确定在特定环境或载体中是否存在可能对人体健康造成不良影响的生物、化学或物理因子的过程。对于饮用水中微生物风险评价,核心是识别相关的病原微生物,并定性描述其可能引起的健康效应(如胃肠炎、死亡)。在水环境中优先关注嗜肺军团菌(淋浴或喷雾吸入)[1]、机会病原微生物、原虫与肠道病毒等病原;必要时将耐药性风险纳入情景。场景界定需要从饮用水系统全过程中选择,明确工况和生物膜等驱动变量[52]。
暴露评价是定量或定性估计目标人群在特定时期内暴露于危害因子的强度、频率和持续时间的全过程;包括对危害因子在环境中的浓度、人群的接触途径和行为模式进行综合评估。暴露评估的量化路径一般由饮用暴露按浓度×摄入量估计;淋浴或喷雾吸入可结合粒径分布和肺沉积模型与活动时长和频次参数,形成日暴露分布。方法与参数可参考美国毒物与疾病登记署(Agency for Toxic Substances and Disease Registry, ATSDR)暴露与剂量评价。军团菌案例中,常将水龄、军团菌浓度、感染或重症概率串联(淋浴情景),评估“降龄、冲洗”等干预的风险下降幅度;不同淋浴气溶胶模型与粒径划分对结果较敏感,需结合设备与现场气溶胶测定[53]。暴露评估的输入来自培养、qPCR或dPCR等分子检测方法,必要时辅以宏基因组识别功能风险[54]。本地基线可参考我国2018-2022年湖北省33 384份集中式供水样本微生物监测[55]。
剂量-反应评价是定量描述人体暴露于某种危害因子的剂量(如摄入的病原微生物数量)与产生不良健康反应(如感染、发病或死亡)的概率之间关系的过程。军团菌一般采用吸入的方式作为剂量反应的模型,最新系统综述或元分析汇总了qPCR与培养之间的换算关系,避免风险高估[11]。诺如病毒以摄入为模型,GII.2人群挑战试验(JID 2022)更新了低剂量端感染或发病概率和宿主易感性的差异参数,为饮水场景外推提供依据[56]。隐孢子虫病使用摄入模型进行计算[57],多项研究从处理效果(凝沉滤)与灵敏度分析校准低剂量风险,支持以LRV和QMRA联用来制定工艺控制[50]。当剂量-反应参数存在不一致或不完备的问题,可参考美国环境保护署2025风险基准框架中的参考病原与剂量-反应目标表。
风险表征的基本原理是将以上3个步骤的结果进行整合与量化不确定性,给出日或年感染风险或伤残调整寿命年,并与健康阈值进行对标,从而为管理决策提供依据。US EPA在2025年发布面向再生或饮用回用的风险基准框架,系统给出剂量-反应参数、伤残调整寿命年对标与对数削减目标的制定方法,便于将风险表征转变为工艺目标。风险表征的核心任务是将浓度和暴露分布与剂量-反应结合,产出可对标(健康阈值或等效年感染风险10-3)的感染概率(probability of infection, P_inf)或伤残调整寿命年指标,以决定是否需要冲洗、提高余氯、温控等措施。降低水龄、有序冲洗和维持残氯等可量化降低感染概率和伤残调整寿命年,但参数独立性假设与相关性处理会影响结果,需开展敏感性分析[12]。
由于数据不完整和模型简化会造成风险评估结果的不确定性,为了使QMRA的结果更准确,对暴露评估和剂量-反应分析进行不确定分析尤为重要。目前多采用随机分析的方法解决不确定性问题,对QMRA的风险进行动态表征。蒙特卡洛模拟[53]通过生成大量的随机数(>10 000个)来近似计算风险值,运用概率密度方法传播参数的不确定性,可以处理复杂模型中的不确定性问题,在风险表征中应用越来越普遍。病原微生物在介质中的浓度、单次暴露时间、暴露频率等均是产生随机数之前需要明确的变量[56]。敏感性分析是识别和排序哪些参数的不确定性对最终风险结果影响最大的关键方法,它常与蒙特卡洛模拟结合使用。在蒙特卡洛模拟的基础上,使用统计方法(如Spearman秩相关系数、回归分析)计算每个输入参数分布与最终风险分布之间的关联强度[12,57-58]。
为便于本土化实施,建议按照“WSP-QMRA”思路并结合微生物检测方法,将风评模型变成可执行流程,如图2所示:先用本地监测数据建立关键输入的本土化分布(病原微生物原水浓度、处理LRV、管网分区差异),再用全局敏感性分析给出“最影响风险的几个因子”,确定关键影响因子与关键控制点,最后将风险分布反推成可执行阈值,联合SOP明确风险控制的触发条件[13];在实际操作中,高频在线检测方法和高分辨率检测更容易形成可追溯的指标:例如全规模online-FCM在水厂进水端观测到细胞浓度波动,能捕捉常规在线指标无法及时反映的水质变化,用于事件预警与生物稳定性追踪[7];宏基因组研究追踪到ARG谱系并给出微生物群落和环境因子间的关联,可作为抗微生物药物耐药性(antimicrobial resistance, AMR)预警与参数本土化的直接数据模板[49],用真实事件做外部校验,从而让模型、异常监测和处置真正闭环并可持续发展。
本文系统梳理了饮用水中病原微生物定量监测与风险评估的核心进展,明确当前领域全过程治理的转变。阐明了病原微生物在配水系统与建筑末梢生物膜中的生存机制与污染风险。在分析传统培养法合规性价值与局限性的基础上,重点评述了与宏基因组学、宏病毒组学相融合的高新技术在危害识别与实时预警中的应用优势。此外,本文总结了利用内标法等策略将微生物相对丰度转化为计算所需的绝对剂量,同时说明了分子检测结果的保守换算原则与不确定性表征方法,为QMRA在应用中由于数据不完整和模型简化所造成风险评估结果的不确定性提供解决思路。
面对日益复杂的环境微生物威胁,未来的饮用水微生物安全管理必须从单一的终端水质达标,向“风险预警-精准防控-健康导向”的全链条智能化治理模式转变,如图3所示。
该未来治理体系框架主要由四大维度构成并形成闭环:(1)底层支撑,强调以环境病原微生物的赋存规律为基础,依托dPCR、宏基因组等现代分子生物学定量检测技术,实现对水环境中病原微生物的高精度追踪与数据累积;(2)核心方法论,即利用系统化的QMRA将病原浓度转化为人群感染概率与疾病负担等直观的健康风险指标;(3)由评估向实践转化的治理体系,未来应深度融合大数据与机器学习,将静态的QMRA转化为动态风险预测,精准锁定对公共健康威胁最大的关键病原体及水处理流程中的关键控制点(critical control points, CCPs);(4)全链条的智能化展望,这也是未来发展的终极形态。通过物联网、AI与在线传感技术的结合,实现“从源头到龙头”的实时智能监测与靶向控制,最终将风险评估结果无缝对接到公共卫生政策制定、水质标准修订及突发公共卫生事件的应急管理中,全面筑牢饮用水生物安全防线。
本文整合国际规范与本土实践,为供水单位提供了场景化监测与参数本地化结合的实施思路,也为监管机构制定政策提供技术支撑,服务于水质指南、基准、标准的制订与修订,提升病原微生物的精准管控能力,切实降低饮用水综合健康风险。
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2026年第66卷第10期
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doi: 10.13343/j.cnki.wsxb.20260172
  • 接收时间:2026-03-03
  • 首发时间:2026-10-13
  • 出版时间:2026-10-04
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  • 收稿日期:2026-03-03
  • 录用日期:2026-03-26
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    1.华南农业大学 食品学院,广东 广州
    2.广东省科学院微生物研究所,华南应用微生物国家重点实验室,广东省微生物安全与健康重点实验室,国家卫健委微生物食品营养与安全科技创新平台,国家市场监督管理总局重点实验室(食品微生物安全 大数据技术),广东 广州
    3.广东环凯信息科技有限公司,广东 广州

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2种不同金属材料的力学参数
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genus
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species
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Percentage of
total species (%)
属
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species
占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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