Article(id=1324078524130272111, tenantId=1146029695717560320, journalId=1227665162245664772, issueId=1324078333767594037, articleNumber=null, orderNo=null, doi=10.20043/j.cnki.MPM.202508014, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1754064000000, receivedDateStr=2025-08-02, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1793614520341, onlineDateStr=2026-11-02, pubDate=1767974400000, pubDateStr=2026-01-10, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1793614520341, onlineIssueDateStr=2026-11-02, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1793614520341, creator=13701087609, updateTime=1793614520341, updator=13701087609, issue=Issue{id=1324078333767594037, tenantId=1146029695717560320, journalId=1227665162245664772, year='2026', volume='53', issue='1', pageStart='1', pageEnd='192', issueExtLink='null', onlineDate='null', pubDate='1767974400000', pubDateStr='2026-01-10', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=0, createTime=1793614474956, creator='13701087609', updateTime=1793614571752, updator='13701087609', preIssue=null, nextIssue=null, articleTotal=null, ext={EN=IssueExt(id=1324078739914621270, tenantId=1146029695717560320, journalId=1227665162245664772, issueId=1324078333767594037, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1324078739914621271, tenantId=1146029695717560320, journalId=1227665162245664772, issueId=1324078333767594037, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null, downloadFileDto=null}, startPage=173, endPage=180, ext={EN=ArticleExt(id=1324078524314821488, articleId=1324078524130272111, tenantId=1146029695717560320, journalId=1227665162245664772, language=EN, title=Etiological surveillance of Vibrio cholerae in Shaanxi Province,2021—2024, columnId=1228016571713520579, journalTitle=Modern Preventive Medicine, columnName=Health Supervison, runingTitle=null, highlight=null, articleAbstract=
Objective

To investigate the distribution and pathogenic characteristics of Vibrio cholerae in aquatic products (such as soft-shelled turtles), aquatic environments and humans in Shaanxi Province.

Methods

Vibrio cholerae strains reported through the National Pathogen Identification Network from 2021 to 2024 were collected. The distribution of strains from humans and environmental sources was described and analyzed. Serogroup identification and virulence gene detection were performed.Core-genome multilocus sequence typing(cgMLST) and core-genome single nucleotide polymorphism (cgSNP) analyses were performed.

Results

A total of 198 Vibrio cholerae strains were collected from 2021 to 2024, including 14 clinical isolates (9 cases of diarrhea and 5 blood samples) and 184 environmental isolates. The overall environmental positivity rate was 34.65% (184/531), with the highest rate in Xi’an City (66.35%), followed by Ankang City (32.66%) and Hanzhong City (21.93%). The positivity rate in sales part was 42.07%, higher than that in farming part (26.92%). The positivity rate in soft-shelled turtle samples was 46.56%, significantly higher than that in other aquatic products (23.77%). Serotyping revealed 3 O1 strains, 13 O139 strains, and the remainder as non-O1/non-O139. Nine strains were positive for the ctxA gene. cgMLST analysis identified 26 MST Clusters, with all ctxA-positive strains clustering in a single lineage: three ST69 strains formed one cluster and five ST168 strains formed another, while the remaining strains were genetically distant. cgSNP analysis of the 9 ctxA-positive strains showed that ST69 and ST168 strains belonged to distinct lineages, differing by 8 100 SNPs, whereas strains of the same ST type exhibited minimal SNP differences, indicating high genetic stability. The patient isolate from Hanzhong City and the soft-shelled turtle isolate from Ankang City differed by only 2 SNPs.

Conclusion

In Shaanxi Province, Vibrio cholerae in the environment poses a relatively high risk at the farming and sales stages of soft-shelled turtles and other aquatic products, and a small proportion of isolates carry the ctxA virulence gene. Core-genome analysis indicates that toxigenic strains are genetically stable and may be indirectly linked to clinical cases. Surveillance and early warning for diarrheal cases and along the soft-shelled turtle-related production and supply chain should be strengthened to ensure the timely prevention and control of outbreaks.

, authors=Yi SHI1, Ya-li CHEN2, Wan-jing LIU3, Yang LUAN4, Zheng ZHANG1, Shuang LIANG1, Lin MA1, Shen LI1, Dong-li LIU1, authorsList=Yi SHI, Ya-li CHEN, Wan-jing LIU, Yang LUAN, Zheng ZHANG, Shuang LIANG, Lin MA, Shen LI, Dong-li LIU, authorCompany=null, correspAuthors=Dong-li LIU, authorNote=null, correspAuthorsNote=null, copyrightStatement=null, copyrightOwner=null, extLink=null, articleAbsUrl=null, sourceXml=null, magXml=null, pdfUrl=null, pdf=null, pdfFileSize=null, pdfExtLink=null, richHtmlUrl=null, mobilePdfUrl=null, reviewReport=null, pdfFirstPage=null, abstractGraph=null, abstractGraphContent=null, abstractVideo=null, citation=null, cebUrl=null, magXmlContent=null, mapNumber=null, fund=null), CN=ArticleExt(id=1324078526982398839, articleId=1324078524130272111, tenantId=1146029695717560320, journalId=1227665162245664772, language=CN, title=陕西省2021—2024年霍乱弧菌病原学监测结果分析, columnId=1228016571923235795, journalTitle=现代预防医学, columnName=卫生监督, runingTitle=null, highlight=null, articleAbstract=
目的

了解陕西省霍乱弧菌在甲鱼等水产品、水体环境及人群中的分布规律及病原学特征。

方法

收集2021—2024年国家致病菌识别网上报的霍乱弧菌菌株,描述分析菌株在人群及环境中的分布规律,对菌株进行血清群鉴定及毒力基因检测,对菌株进行核心基因组多位点序列分析(core genome multi-locus sequence analysis,cgMLST)及核心基因组单核苷酸多态性分析(core genome single nucleotide polymorphism analysis,cgSNP)分析。

结果

2021—2024年共收到菌株198株,其中分离自病人菌株14株(腹泻类样本9株,血液样本5株),环境分离菌株184株。环境样本中霍乱弧菌分离阳性率为34.65%(184/531),西安市阳性率最高为66.35%,其次安康市为32.66%,汉中市为21.93%;销售环节阳性率42.07%,高于养殖环节阳性率的26.92%;甲鱼的阳性率为46.56%,明显高于其他水产品的23.77%。霍乱弧菌血清分型O1群3株,O139群13株,其余菌株为非O1非O139群,CTX阳性菌株9株。cgMLST分析共有26个MST Cluster,CTX基因阳性的霍乱弧菌聚在一个分支,其中有3株ST69型聚为一簇,ST168型5株聚为一簇,其余菌株距离较远。对CTX阳性的9株霍乱弧菌进行cgSNP分析,ST69与ST168分属于不同分支之间,相差8 100个SNPs,同ST型的菌株SNP差异很小,分离自汉中市的病人样本与分离自安康市的甲鱼样本,相差2个SNPs。

结论

陕西省环境中霍乱弧菌在甲鱼等水产品的养殖和销售环节风险较高,少数菌株有CTX毒力基因,核心基因组分析霍乱弧菌产毒株遗传变异稳定,且与病例存在间接的关联。应加强腹泻病例及甲鱼相关产业链条的监测预警,及时预防控制疫情发生。

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石一(1981—),男,硕士,副主任技师,研究方向:病原微生物

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刘东立,E-mail:
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注:图中包含所有acf+菌株18株(含ctx+菌株9株,ctx-菌株9株),ctx-/acf-菌株随机选择18株。

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注:图中栯圆圈中的为CTX阳性菌株,红色虚圈为人源菌株。

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Detection of Vibrio cholerae strains in environmental samples in Shaanxi Province

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地市检测数阳性数阳性率(%)χ2值P值
西安1046966.3562.780<0.001
汉中2285021.93
安康1996532.66
样本类别
环境水体等2789433.810.1120.738
水产品2539035.57
传播环节
销售27111442.0712.778<0.001
养殖2607026.92
合计53118434.65
), ArticleFig(id=1324078532678263741, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1324078524130272111, language=CN, label=表1, caption=

陕西省环境样本中霍乱菌株检出情况

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地市检测数阳性数阳性率(%)χ2值P值
西安1046966.3562.780<0.001
汉中2285021.93
安康1996532.66
样本类别
环境水体等2789433.810.1120.738
水产品2539035.57
传播环节
销售27111442.0712.778<0.001
养殖2607026.92
合计53118434.65
), ArticleFig(id=1324078532766344126, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1324078524130272111, language=EN, label=Table 2, caption=

Positive detection rates of Vibrio cholerae in different water samples

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样本类别检测数阳性数阳性率(%)χ2值P值
自然水体1024039.221.7370.187
养殖水体1765430.68
合计2789433.81
), ArticleFig(id=1324078532850230207, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1324078524130272111, language=CN, label=表2, caption=

不同水体样本霍乱弧菌检出阳性率情况

, figureFileSmall=null, figureFileBig=null, tableContent=
样本类别检测数阳性数阳性率(%)χ2值P值
自然水体1024039.221.7370.187
养殖水体1765430.68
合计2789433.81
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Detection rates of Vibrio cholerae in turtle and other aquatic products samples

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样本类别检测数阳性数阳性率(%)χ2值P值
甲鱼1316146.5613.344<0.001
其他水产1222923.77
合计2539035.57
), ArticleFig(id=1324078532997030849, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1324078524130272111, language=CN, label=表3, caption=

甲鱼与其他水产品样本霍乱弧菌检出阳性率情况

, figureFileSmall=null, figureFileBig=null, tableContent=
样本类别检测数阳性数阳性率(%)χ2值P值
甲鱼1316146.5613.344<0.001
其他水产1222923.77
合计2539035.57
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陕西省2021—2024年霍乱弧菌病原学监测结果分析
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石一 1 , 陈雅丽 2 , 刘万静 3 , 栾阳 4 , 张铮 1 , 梁爽 1 , 马琳 1 , 李慎 1 , 刘东立 1
现代预防医学 | 卫生监督 2026,53(1): 173-180
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现代预防医学 |卫生监督 2026 , 53 (1) : 173 -180
陕西省2021—2024年霍乱弧菌病原学监测结果分析
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石一1, 陈雅丽2, 刘万静3, 栾阳4, 张铮1, 梁爽1, 马琳1, 李慎1, 刘东立1
作者信息
  • 1.陕西省疾病预防控制中心病原微生物与生物检验所,陕西 西安 710054
  • 2.汉中市疾病预防控制中心检验科
  • 3.安康市疾病预防控制中心生物检验科
  • 4.西安市疾病预防控制中心微生物与生物检验科
通讯作者:
刘东立,E-mail:
作者简介:

石一(1981—),男,硕士,副主任技师,研究方向:病原微生物

Etiological surveillance of Vibrio cholerae in Shaanxi Province,2021—2024
Yi SHI1, Ya-li CHEN2, Wan-jing LIU3, Yang LUAN4, Zheng ZHANG1, Shuang LIANG1, Lin MA1, Shen LI1, Dong-li LIU1
Affiliations
  • Institute of Pathogenic Microbiology and Biological Testing, Shaanxi Provincial Center for Disease Control and Prevention, Xi’an, Shaanxi 710054, China
出版时间: 2026-01-10 doi: 10.20043/j.cnki.MPM.202508014
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目的

了解陕西省霍乱弧菌在甲鱼等水产品、水体环境及人群中的分布规律及病原学特征。

方法

收集2021—2024年国家致病菌识别网上报的霍乱弧菌菌株,描述分析菌株在人群及环境中的分布规律,对菌株进行血清群鉴定及毒力基因检测,对菌株进行核心基因组多位点序列分析(core genome multi-locus sequence analysis,cgMLST)及核心基因组单核苷酸多态性分析(core genome single nucleotide polymorphism analysis,cgSNP)分析。

结果

2021—2024年共收到菌株198株,其中分离自病人菌株14株(腹泻类样本9株,血液样本5株),环境分离菌株184株。环境样本中霍乱弧菌分离阳性率为34.65%(184/531),西安市阳性率最高为66.35%,其次安康市为32.66%,汉中市为21.93%;销售环节阳性率42.07%,高于养殖环节阳性率的26.92%;甲鱼的阳性率为46.56%,明显高于其他水产品的23.77%。霍乱弧菌血清分型O1群3株,O139群13株,其余菌株为非O1非O139群,CTX阳性菌株9株。cgMLST分析共有26个MST Cluster,CTX基因阳性的霍乱弧菌聚在一个分支,其中有3株ST69型聚为一簇,ST168型5株聚为一簇,其余菌株距离较远。对CTX阳性的9株霍乱弧菌进行cgSNP分析,ST69与ST168分属于不同分支之间,相差8 100个SNPs,同ST型的菌株SNP差异很小,分离自汉中市的病人样本与分离自安康市的甲鱼样本,相差2个SNPs。

结论

陕西省环境中霍乱弧菌在甲鱼等水产品的养殖和销售环节风险较高,少数菌株有CTX毒力基因,核心基因组分析霍乱弧菌产毒株遗传变异稳定,且与病例存在间接的关联。应加强腹泻病例及甲鱼相关产业链条的监测预警,及时预防控制疫情发生。

霍乱弧菌  /  霍乱肠毒素  /  cgMLST  /  cgSNP
Objective

To investigate the distribution and pathogenic characteristics of Vibrio cholerae in aquatic products (such as soft-shelled turtles), aquatic environments and humans in Shaanxi Province.

Methods

Vibrio cholerae strains reported through the National Pathogen Identification Network from 2021 to 2024 were collected. The distribution of strains from humans and environmental sources was described and analyzed. Serogroup identification and virulence gene detection were performed.Core-genome multilocus sequence typing(cgMLST) and core-genome single nucleotide polymorphism (cgSNP) analyses were performed.

Results

A total of 198 Vibrio cholerae strains were collected from 2021 to 2024, including 14 clinical isolates (9 cases of diarrhea and 5 blood samples) and 184 environmental isolates. The overall environmental positivity rate was 34.65% (184/531), with the highest rate in Xi’an City (66.35%), followed by Ankang City (32.66%) and Hanzhong City (21.93%). The positivity rate in sales part was 42.07%, higher than that in farming part (26.92%). The positivity rate in soft-shelled turtle samples was 46.56%, significantly higher than that in other aquatic products (23.77%). Serotyping revealed 3 O1 strains, 13 O139 strains, and the remainder as non-O1/non-O139. Nine strains were positive for the ctxA gene. cgMLST analysis identified 26 MST Clusters, with all ctxA-positive strains clustering in a single lineage: three ST69 strains formed one cluster and five ST168 strains formed another, while the remaining strains were genetically distant. cgSNP analysis of the 9 ctxA-positive strains showed that ST69 and ST168 strains belonged to distinct lineages, differing by 8 100 SNPs, whereas strains of the same ST type exhibited minimal SNP differences, indicating high genetic stability. The patient isolate from Hanzhong City and the soft-shelled turtle isolate from Ankang City differed by only 2 SNPs.

Conclusion

In Shaanxi Province, Vibrio cholerae in the environment poses a relatively high risk at the farming and sales stages of soft-shelled turtles and other aquatic products, and a small proportion of isolates carry the ctxA virulence gene. Core-genome analysis indicates that toxigenic strains are genetically stable and may be indirectly linked to clinical cases. Surveillance and early warning for diarrheal cases and along the soft-shelled turtle-related production and supply chain should be strengthened to ensure the timely prevention and control of outbreaks.

Vibrio cholerae  /  Cholera toxin  /  cgMLST  /  cgSNP
石一, 陈雅丽, 刘万静, 栾阳, 张铮, 梁爽, 马琳, 李慎, 刘东立. 陕西省2021—2024年霍乱弧菌病原学监测结果分析. 现代预防医学, 2026 , 53 (1) : 173 -180 . DOI: 10.20043/j.cnki.MPM.202508014
Yi SHI, Ya-li CHEN, Wan-jing LIU, Yang LUAN, Zheng ZHANG, Shuang LIANG, Lin MA, Shen LI, Dong-li LIU. Etiological surveillance of Vibrio cholerae in Shaanxi Province,2021—2024[J]. Modern Preventive Medicine, 2026 , 53 (1) : 173 -180 . DOI: 10.20043/j.cnki.MPM.202508014
霍乱是由霍乱弧菌引起的急性、烈性肠道传染病,被世界卫生组织列为国际检疫传染病,也是中国《中华人民共和国传染病防治法》规定的甲类传染病之一。霍乱弧菌广泛分布于自然环境的水体和水产品中,O1 群和 O139 群能引发霍乱的流行和大流行。自 2021 以来,霍乱病例数有所增加,在全球多个国家分布扩散,报告病例数和病死率均有上升[1]。在2015—2024年,国内霍乱保持在低流行水平,平均每年报告18例,且食源传播已为主要模式[2]。历史上陕西省曾有霍乱流行,自20世纪80年代后再无病例报道。2022年陕西省城固县报告一起霍乱散发疫情,是本省40余年来首次报告的霍乱病例[3]。为了解陕西省环境及人群中霍乱弧菌传播状况,2023年开始陕西省在汉中、安康、西安市针对环境水体、水产品养殖及销售环节分离霍乱弧菌,收集2021年以来临床分离菌株,开展霍乱弧菌发生与传播因素监测,本文对监测收集到的菌株数据进行分析,揭示其在人群及环境中的分布规律,探索菌株的血清群及毒力基因特征。
收集 2021年 1 月 1 日— 2024年12 月 31 日国家致病菌识别网上报的陕西省霍乱弧菌菌株,环境监测选择秦岭南部的汉中、安康市以及省会西安市。详细记录菌株的分离地点、分离时间、样本类型(病人样本或环境样本,若为病人样本,记录样本是腹泻类样本还是血液样本;若为环境样本,记录是环境水样、水产品样本,以及样本采集的环节是养殖环节还是销售环节等信息)。
采用玻片凝集试验及荧光定量聚合酶链式反应(polymerase chain reaction,PCR)方法对分离的霍乱弧菌菌株进行血清群鉴定,使用标准的 O1 群和 O139 群霍乱弧菌抗血清(日本生研)及霍乱弧菌/O1群/O139群/CTX基因多重荧光定量PCR试剂盒(深圳生科原),按照试剂盒说明书进行操作。
使用细菌基因组 DNA 提取试剂盒(西安天隆)提取霍乱弧菌菌株的基因组 DNA,采用《国家致病菌识别网技术手册》推荐的方案,使用Nextera XT DNA Library Preparation Kit建库试剂盒(北京微未来科技有限公司)构建测序文库,选择MiniSeq测序平台(美国Illumina公司),使用MiniSeq High Output Kit (300 cycles)(北京微未来科技有限公司)测序试剂盒进行上机测序,测序策略为PE150,测序深度不低于 100×,高质量测序数据(Q20)≥1Gb,Q30比例≥85%,利用国家致病菌识别网终端对测序数据进行质量控制和拼接,基因组覆盖度≥95%,>1 000 bp的Scaffold数目小于100。
菌株测序数据导入Riodom Seqphere 10.0 5软件[4],利用集成的V.cholerae VFDB[5]预测并查询菌株基因组携带的毒力基因。使用Chiplot(https://www.chiplot.online/)绘制不同样本的毒力基因热图。
采用RiodomSeqphere 10.0 5软件[3]提供的V. cholerae cgMLST 3 251 targets方案,对良好等位基因位点(perc. Good target)在80%以上的菌株进行cgMLST分型分析,构建最小生成树图(minimum spanning tree,MST),MST Cluster阈值设置为20个等位基因,分析霍乱弧菌菌株之间的遗传关系。
对携带霍乱肠毒素(cholera enterotoxin,CTX)毒力基因的菌株,用RiodomSeqphere 10.0 5软件搜寻核心基因组单核苷酸变异位点,绘制系统发育进化树,进行cgSNP分析,评估霍乱弧菌毒力株之间的基因变异程度。
使用 WPS软件对收集到的数据进行整理和描述性统计分析。计算不同样本类型、不同地区菌株的分离率,采用χ2检验比较不同地区、不同样本类型菌株分离率的差异,检验水准α=0.05。
2021—2024年共收到 198 株霍乱弧菌菌株,其中分离自病人的菌株 14 株,分离自环境的菌株 184 株。病人分离株中,9株来自腹泻类样本,5 株来自血液样本;安康市1株,西安市10株,延安市1株,汉中市2株(霍乱病例)。不同地区的环境分离率存在差异(P<0.001),西安市阳性率最高,为66.35%(69/104);其次是安康市,阳性率为 32.66%(65/199);汉中市阳性率为 21.93%(50/228)。见表1。
在环境样本中,环境水样的阳性率为 33.81%(137/405),水产品样本的阳性率为 35.57%(47/126)。在水产品样本中,甲鱼的检出率最高,达到 46.56%(37/79),明显高于其他水产品的23.77%(10/47)。从样本采集环节来看,销售环节样本的阳性率为42.07%(79/188),高于养殖环节的 26.92%(36/134),江河等自然水体与养殖水、不同年份以及环境样本与水产品检出率之间无差异。见表2、3。
对198株霍乱弧菌进行玻片凝集血清分型及O1/O139/CTX多重荧光定量PCR检测,其中O1群3株,CTX毒力基因阴性,O139群13株,有3株CTX毒力基因阳性。非O1非O139群181株,有6株CTX毒力基因阳性,环境及水产品样本CTX阳性率为3.26%。
通过VFDB数据库查询预测共有毒力基因8类48种4 690个。所有菌株均有hlyA(霍乱弧菌细胞毒素)、lusX(S-核糖基高半胱氨酸酶)及vasF(VI型分泌系统蛋白VasF),大部分菌株均有VI型分泌系统的相关毒力基因如:vas(192)、vgrG(46)、VCA(190)等,RTX毒素相关毒力基因如rtxC(187)、rtxD(187)、rtxB(185);有18株菌携带有acf(辅助定植因子)及毒素共调节菌毛蛋白(tcp)相关毒力基因,占9.09%(18/198),有9株携带有CTX(霍乱肠毒素)、ace(辅助霍乱肠毒素)等毒力基因,占 4.55%(9/198)。见图1。
携带CTX毒力基因的菌株有2株来自病例,4株来自甲鱼,2株来自养殖水体,1株来自其他水产品(多宝鱼)。环境及水产品样本中CTX毒力基因检出率为3.80%(7/184),CTX阳性的菌株同时也有较多的其他毒力基因,例如来自汉中市2022年霍乱病例菌株,携带41个毒力基因,其次是2023年安康市的甲鱼分离菌株,有40个毒力基因,汉中市2022年病例密切接触者的菌株有39个毒力基因,另外2024年西安市、汉中市甲鱼分离菌株均携带了37~39个毒力基因。
不同样本类别中来自病人样本的菌株携带毒力基因最多,平均26.85个(349/13),其次为甲鱼样本,平均26.00个(1 352/52),其他水产品平均为23.85个(954/40),养殖水体为23.29个(955/41),最小的为自然水体的22.98个(1 080/47)。不同样本间毒力基因数量分布具有差异性(F=2.44,P<0.05)。
全基因组测序数据分析对霍乱弧菌进行了传统的MLST型,共分型成功49个ST型,其中ST168最多,有6株,ST1600、ST689、ST846各4株,ST69、ST718、ST853、ST886各3株,ST1160、ST1213、ST1310、ST1317、ST1723、ST271、ST334各2株,有1株的ST型有33株,另外还有117株未分型。
筛选核心基因组良好的等位基因位点80%以上的菌株共176株进行cgMLST分析,发现CTX阳性的菌株cgMLST位点覆盖在90%以上。绘制MST图聚类结果显示,以20个等位基因为距离计算,共有26个MST Cluster,携带毒力基因的霍乱弧菌聚在一个分支,其中有3株O139群ST69型聚为一簇,2株分离自汉中市的病人样本,另一株分离自安康市甲鱼样本,相差2个等位基因,ST168型5株聚为一簇,等位基因相差4~16个,均分离自甲鱼相关的样本。另一株O139群分离自多宝鱼样本的菌株相差等位基因33个,与人体样本之间相差1 282个等位基因。其余无毒力基因的菌株之间变异很大,无论是O1群、O139群和非O1非O139群,相差多在2 000个等位基因左右。其余的Cluster中的菌株,多为同一时段分离自同一地点相同的样本。人群分离的非O1非O139且CTX(-)菌株与环境菌株无明显关联,2023年雁塔区2株腹泻病例较接近,其余菌株均较分散。见图2。
对CTX阳性的9株霍乱弧菌进行cgSNP分析,共查找到ST69与ST168分属于不同分支之间相差8 100个SNPs,2株分离自汉中市的病人样本,另一株分离自安康市有甲鱼样本,相差2个SNPs。另外6株分离自甲鱼相关样本的ST168型,菌株之间相差4~13个SNPs,同一时间同一地点分离菌株SNP无差异。见图3。
自2022年起,陕西省逐步实施了霍乱弧菌监测项目,重点覆盖了陕南地区的汉中市、安康市以及省会西安市等关键区域。监测活动包括采集各类水体和水产品的样本,并收集了部分临床病例中的霍乱分离菌株,首次对陕西省霍乱弧菌的分布和病原学特征进行了深入研究。监测结果揭示了霍乱弧菌在环境中的检出率为34.65%,这一比例高于国内广州[6]、梧州市[7]、赣州市[8]的报告数据,与广东省清远市近年来的监测结果相近[9],这一趋势与近年来霍乱疫情的上升趋势相吻合。西安市的阳性检出率显著高于汉中市和安康市,主要由于西安市的采样地点均为大型海鲜水产批发市场,样本类型繁多且市场流通量大,从而增加了霍乱弧菌的传播风险。相比之下,汉中市和安康市的采样点主要集中在小型养殖水体、自然水体或小型市场,因此检出率相对较低。本研究的监测数据表明,在销售环节的阳性率明显高于养殖环节,可能与销售环节为来自各地水产品的集散地,流通交易频繁,环境卫生较养殖环节差有关。虽然环境样本霍乱弧菌阳性率较高,但同期临床感染霍乱病例并不多,一方面近年来我国用水安全卫生极大改善,像霍乱、细菌性痢疾等介水传播疾病传播风险大幅下降,另一方面临床感染的菌株多为CTX(-)的弱毒株,国内也有临床分离出非O1非O139群且无毒力基因的菌株[10-11],这类霍乱弧菌血流感染事件往往与典型霍乱无关[12]。本研究临床病例中有5株血液分离株及7株腹泻样本分离株,均为CTX(-)的非O1非O139菌株,与环境及水体监测的多数菌株一致,对人群致病力较低,流行传播风险较小。
近年来国内典型的霍乱疫情多通过甲鱼等食物散发传播[13-14]。本研究水产品样本中,甲鱼的阳性率显著高于其他水产品,这与广东省[9]等12个省市[15]的报道一致。且携带CTX毒力基因的菌株以甲鱼样本最为突出,这些菌株有可能引起典型的严重霍乱腹泻症状。提示我们在养殖、流通销售环节以及甲鱼是霍乱弧菌传播的关键环节。在本研究中,还发现3株血清群为O1的菌株和10株O139群的菌株,但未携带CTX毒力基因,属O1或O139群非产毒菌株,其致病力有限,症状多不典型[16-17]。近年来,国内外有报道非O1非O139型菌株携带CTX毒力基因并引起霍乱的情况[18],本研究也发现有6株CTX(+)的非O1非O139群菌株,此类菌株可能具有较强的致病潜力及传播风险,应予以关注。
霍乱弧菌的基因组中含有由丝状溶原性噬菌体CTXφ编码的霍乱肠毒素基因ctxAB[19],以及tcp-acf基因簇、整合酶基因、转位酶基因,这些连同两侧的att序列共同构成了霍乱弧菌的毒力岛[20]。值得注意的是,尽管某些菌株的CTX检测结果为阴性,但若它们携带毒力岛,同样需要引起我们的重视。本研究的数据显示,9株CTX基因阳性的菌株均含有由acf及tcp相关基因构成的毒力岛(ctx+/acf+),并且这些菌株往往携带更多的其他毒力基因,为典型的霍乱强毒株,具有较强的致病性。相比之下,大多数CTX阴性的菌株并不包含毒力岛的相关基因(ctx-/acf-),一般认为致病性较弱。然而,本研究中还发现9株CTX基因阴性但具有毒力岛结构的菌株(ctx-/acf+),它们的毒力基因表达受到抑制,导致致病性显著降低[21]。据报告,水产毒株有可能将溶原性噬菌体转移给这些菌株,使它们重新获得致病力[22]。因此,那些携带有毒力岛tcp-acf基因簇的菌株可能成为潜在的毒力菌株,应加强基因组水平的监测并研究其毒力因子传递的规律。
通过对霍乱弧菌核心基因组的遗传进化和变异进行分析,可以实现分子溯源和进化关系的深入研究,这对于霍乱的防控具有至关重要的意义[23]。本研究采用cgMLST分析方法揭示,CTX基因阴性的菌株之间存在较大变异,不同菌株间的等位基因差异可高达上千个,而CTX阳性的菌株则相对稳定,这表明毒力基因的存在可能对菌株的遗传稳定性产生影响。CTX阳性的9株霍乱弧菌基因组显著大于CTX阴性的菌株,且cgMLST核心位点的覆盖度均超过90%,这说明遗传稳定性与毒力基因岛的完整性密切相关。进一步的cgSNP分析显示,CTX阴性的菌株间SNP位点差异高达数万个,而CTX阳性基因的菌株差异较小,除了相同序列类型(ST)的菌株外,大多数菌株之间并不紧密相关[16]。本研究构建的系统发育树显示,ST69型与ST168型之间的SNPs差异为8 100个,而相同ST型内的差异不超过13个。ST69型是近年来国内流行的菌株[24],2022年在汉中市城固县发生的O139群霍乱疫情中,ST69型菌株被发现,当年未能追溯到源头[1]。然而,2023年在安康市某水产品市场销售的甲鱼中检测出CTX阳性的O139群菌株,与临床病例中的菌株仅相差2个SNPs,这表明这些菌株之间可能存在某种潜在的关联性,但在时间序列上存在矛盾。有可能该毒株在国内,随着甲鱼等水产品隐匿传播[25],需要更多的样本及流行病学证据来证实。而人群分离的非O1非O139菌株之间无关联,与环境菌株遗传距离也较远无关联,与弧菌属基因组变异较大有关。
对陕西省霍乱弧菌的病原学特征进行分析后发现,省内水体及养殖、销售环节中均存在霍乱弧菌的检出情况。特别是与甲鱼相关的链条样本中,CTX阳性菌株的检出率较高,且与病例有一定关联,这提示我们需要关注其潜在的传播风险。本研究仅覆盖了三个地市的霍乱弧菌监测,后续可扩大监测范围,重点关注CTX毒力基因。加强甲鱼养殖环境的监测与消毒,提高公众的卫生意识,加强肠道传染病防控知识的宣传,确保生熟食物分开处理,从而降低霍乱疫情的发生概率。
  • 陕西省重点研发计划(2025SF-YBXM-234)
  • 陕西省疾控局公共卫生科研创新团队(202503)
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2026年第53卷第1期
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doi: 10.20043/j.cnki.MPM.202508014
  • 接收时间:2025-08-02
  • 首发时间:2026-11-02
  • 出版时间:2026-01-10
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  • 收稿日期:2025-08-02
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陕西省重点研发计划(2025SF-YBXM-234)
陕西省疾控局公共卫生科研创新团队(202503)
作者信息
    1.陕西省疾病预防控制中心病原微生物与生物检验所,陕西 西安 710054
    2.汉中市疾病预防控制中心检验科
    3.安康市疾病预防控制中心生物检验科
    4.西安市疾病预防控制中心微生物与生物检验科

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2种不同金属材料的力学参数
科
Family
属数
Number of
genus
种数
Number of
species
占总种数比例
Percentage of
total species (%)
属
Genus
种数
Number of
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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