Article(id=1324078309109269161, tenantId=1146029695717560320, journalId=1227665162245664772, issueId=1324078175000605624, articleNumber=null, orderNo=null, doi=10.20043/j.cnki.MPM.202508323, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1755792000000, receivedDateStr=2025-08-22, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1793614469076, onlineDateStr=2026-11-02, pubDate=1769270400000, pubDateStr=2026-01-25, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1793614469076, onlineIssueDateStr=2026-11-02, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1793614469076, creator=13701087609, updateTime=1793614469076, updator=13701087609, issue=Issue{id=1324078175000605624, tenantId=1146029695717560320, journalId=1227665162245664772, year='2026', volume='53', issue='2', pageStart='193', pageEnd='384', issueExtLink='null', onlineDate='null', pubDate='1769270400000', pubDateStr='2026-01-25', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=0, createTime=1793614437103, creator='13701087609', updateTime=1793614526046, updator='13701087609', preIssue=null, nextIssue=null, articleTotal=null, ext={EN=IssueExt(id=1324078548130082942, tenantId=1146029695717560320, journalId=1227665162245664772, issueId=1324078175000605624, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1324078548130082943, tenantId=1146029695717560320, journalId=1227665162245664772, issueId=1324078175000605624, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null, downloadFileDto=null}, startPage=252, endPage=258, ext={EN=ArticleExt(id=1324078309352538794, articleId=1324078309109269161, tenantId=1146029695717560320, journalId=1227665162245664772, language=EN, title=Study on the virulence and drug-resistant characteristics of Klebsiella pneumoniae in foods sold,Guizhou, columnId=1228016572783063333, journalTitle=Modern Preventive Medicine, columnName=Nutrition and Food Hygiene, runingTitle=null, highlight=null, articleAbstract=
Objective

To investigate the molecular typing, virulence and antimicrobial resistance characteristics of food-borne Klebsiella pneumoniae in Guizhou Province, providing scientific evidence for targeted food safety risk assessment and regulatory decision-making in our province.

Methods

A total of 38 strains of Klebsiella pneumoniae were isolated from pre-packaged chilled ready-to-eat food and pig livers in 9 cities (states) of Guizhou Province from 2023 to 2024. Strains were screened by string test, whole genome sequencing and antimicrobial susceptibility test.

Results

Four hypermucoviscous Klebsiella pneumoniae strains were isolated from 38 isolates. Through whole genome sequence analysis, 20 KL types, 30 ST types, 80 virulence genes and 35 drug resistance genes were obtained. ST 37 was the main strain. HmKp of ST65-KL2 carried iucA, iroB, ybtS, rmpA and clb genes, while ST490-KL3 carried iucA, iroB, ybtS and allS genes. The main β-lactamase resistance genes were blaSHV-11, blaSHV-1, blaLAP-2. 5 of the strains carried blaOXA-10 gene. The results of antibiotic susceptibility tests showed that the top five resistant antibiotics were ampicillin, tetracycline, chloramphenicol, florfenicol and sulfamethoxazole/trimethoprim, with resistance rates of 81.58%, 50.0%, 44.74%, 44.74% and 28.95% respectively. All isolates were sensitive to carbapenems.

Conclusion

The prevalence of virulence genes in Klebsiella pneumoniae in food in Guizhou Province is high, with a strong pathogenic potencial. The ST types and KL types of the strains are diverse.

, authors=Xin-xu LUO, Jing-zhu XIANG, De-zhu ZHANG, Qian ZHOU, Qin-ni ZHENG, Li ZHOU, authorsList=Xin-xu LUO, Jing-zhu XIANG, De-zhu ZHANG, Qian ZHOU, Qin-ni ZHENG, Li ZHOU, authorCompany=null, correspAuthors=Li ZHOU, 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=1324078310434669233, articleId=1324078309109269161, tenantId=1146029695717560320, journalId=1227665162245664772, language=CN, title=贵州省食品中肺炎克雷伯菌毒力与耐药特征研究, columnId=1228016573034721597, journalTitle=现代预防医学, columnName=营养与食品卫生, runingTitle=null, highlight=null, articleAbstract=
目的

了解贵州省市售食品中肺炎克雷伯菌分子分型、毒力与耐药特征,为我省有针对性的开展食品安全风险评估和监管决策提供科学依据。

方法

2023—2024年贵州省9个市(州)从市售预包装冷藏即食食品、生猪肝中分离的38株肺炎克雷伯菌进行拉丝试验、全基因组测序以及抗生素敏感性试验。

结果

38株分离株共筛选出4株高黏液型肺炎克雷伯菌(hypermucoviscous Klebsiella pneumoniae,hmKp)。通过全基因组序列分析,共获得20种荚膜型(capsule type,KL)、30种序列型(sequence typing,ST)、80种毒力基因和35种耐药基因。ST 37为主要菌株,分子型别为ST65-KL2的hmKp同时携带iucA、iroB、ybtS、rmpA、clb基因,ST490-KL3 肺炎克雷伯菌同时携带iucA、iroB、ybtS、allS基因。β-内酰胺酶类耐药基因以blaSHV-11、blaSHV-1、blaLAP-2为主,5株携带blaOXA-10基因。抗生素敏感性试验结果显示,耐药前五位依次是氨苄西林、四环素、氯霉素、氟苯尼考、磺胺甲噁唑/甲氧苄啶,耐药率分别为81.58%、50.0%、44.74%、44.74%和28.95%,对碳青霉烯类药物均敏感。

结论

贵州省市售食品中肺炎克雷伯菌毒力基因携带率高,致病力强,ST型和KL型呈现多样性。

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罗信旭(1990—),女,硕士,主管技师,研究方向:微生物学

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周藜,E-mail:
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Animal, 2024, 18(11): 101345., articleTitle=Review: antimicrobial resistance of klebsiella pneumoniae isolated from poultry, cattle and pigs, refAbstract=null)], funds=[Fund(id=1324078318932329191, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1324078309109269161, awardId=黔科合支撑[2021]一般435号, language=CN, fundingSource=贵州省科技支撑计划项目(黔科合支撑[2021]一般435号), fundOrder=null, country=null), Fund(id=1324078319007826664, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1324078309109269161, awardId=gzwkj2025-513, language=CN, fundingSource=贵州省卫生健康委科学技术基金项目(gzwkj2025-513), fundOrder=null, country=null), Fund(id=1324078319129461481, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1324078309109269161, awardId=ZDSYS【2023】004, language=CN, fundingSource=贵州省微生物组与传染性疾病防控重点实验室(ZDSYS【2023】004), fundOrder=null, country=null), Fund(id=1324078319301427946, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1324078309109269161, awardId=RCJD2105, language=CN, fundingSource=贵州省传染病预防与控制人才基地科研团队(RCJD2105), fundOrder=null, country=null)], companyList=[AuthorCompany(id=1324078310682133170, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1324078309109269161, xref=null, ext=[AuthorCompanyExt(id=1324078310703104691, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1324078309109269161, companyId=1324078310682133170, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=Guizhou provincial Center for Disease Control and Prevention, Guiyang, Guizhou 550004, China), AuthorCompanyExt(id=1324078310715687604, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1324078309109269161, companyId=1324078310682133170, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=贵州省疾病预防控制中心,贵州 贵阳 550004)])], figs=[ArticleFig(id=1324078317535625947, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1324078309109269161, language=EN, label=Fig.1, caption=Phylogenetic tree of SNP, figureFileSmall=ckDagaff2FZrbfFdIgoM4g==, figureFileBig=82vyAYLynNfFbq/0D/wGrw==, tableContent=null), ArticleFig(id=1324078317627900636, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1324078309109269161, language=CN, label=图1, caption=SNP系统发育树, figureFileSmall=ckDagaff2FZrbfFdIgoM4g==, figureFileBig=82vyAYLynNfFbq/0D/wGrw==, tableContent=null), ArticleFig(id=1324078317887947485, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1324078309109269161, language=EN, label=Fig.2, caption=Virulence gene profile, figureFileSmall=w0u85i+nZNqiS6UJOAPV7Q==, figureFileBig=je4eAqB/3V0rFWtuFzlArA==, tableContent=null), ArticleFig(id=1324078317976027870, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1324078309109269161, language=CN, label=图2, caption=毒力基因谱, figureFileSmall=w0u85i+nZNqiS6UJOAPV7Q==, figureFileBig=je4eAqB/3V0rFWtuFzlArA==, tableContent=null), ArticleFig(id=1324078318047331039, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1324078309109269161, language=EN, label=Fig.3, caption=Antibiotic resistance genes and spectrum of 38 strains of Klebsiella pneumoniae, figureFileSmall=lue9mERRT2ffgXbiSHq4DQ==, figureFileBig=gwOdcynD6u31Tc+3+P54QA==, tableContent=null), ArticleFig(id=1324078318127022816, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1324078309109269161, language=CN, label=图3, caption=38株肺炎克雷伯菌耐药基因与耐药谱图, figureFileSmall=lue9mERRT2ffgXbiSHq4DQ==, figureFileBig=gwOdcynD6u31Tc+3+P54QA==, tableContent=null), ArticleFig(id=1324078318206714593, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1324078309109269161, language=EN, label=Table 1, caption=

Virulence gene detection results

, figureFileSmall=null, figureFileBig=null, tableContent=
毒力种类毒力因子基因名称(%)
细胞黏附Ⅲ型菌毛(Mrk)mrkA (97.37)、mrkB (97.37)、mrkC (97.37)、mrkD (97.37)、mrkF (97.37)、mrkH (97.37)、mrkI (94.74)、mrkJ (97.37)
Ⅰ型菌毛(Fim)fimA (100)、fimB (94.74)、fimC (100)、fimD (100)、fimE (100)、fimF (100)、fimG (100)、fimH (100)、fimI (100)、fimK (92.11)
外排泵ArcABarcA (100)、arcB (100)
铁摄取系统气杆菌素(Aer)iucA (42.11)、iucB (42.11)、iucC (42.11)、iucD (42.11)、iutA (100)
肠杆菌素(Ent)entA (100)、entB (100)、entC (100)、entD (100)、entE (100)、entF (100)、entS (100)、fepA (100)、fepB (100)、fepC (100)、fepD (100)、fepG (100)、fes (100)
沙门菌素(Sal)iroB (7.89)、iroC (5.26)、iroD (7.89)、iroE (100)、iroN (94.74)
耶尔森杆菌素(Ybt)fyuA (10.53)、irp1 (10.53)、irp2 (10.53)、ybtA (10.53)、ybtE (10.53)、ybtP (10.53)、ybtQ (10.53)、ybtS (10.53)、ybtT (10.53)、ybtU (10.53)、ybtX (10.53)
营养因子Allantoin utilizationallA (2.63)、allB (5.26)、allC (5.26)、allD (5.26)、allR (2.63)、allS (5.26)
黏液表型调控基因RcsAB、RmpArcsA (97.37)、rcsB (100)、rmpA (2.63)
毒素ColibactinclbA (2.63)、clbB (2.63)、clbC (0)、clbD (0)、clbE (2.63)、clbF (2.63)、clbG (2.63)、clbH (2.63)、clbI (2.63)、clbJ (2.63)、clbK (2.63)、clbL (2.63)、clbM (2.63)、clbN (2.63)、clbO (2.63)、clbP (2.63)、clbQ(2.63)、clbS (0)
), ArticleFig(id=1324078318311572194, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1324078309109269161, language=CN, label=表1, caption=

毒力基因检出结果

, figureFileSmall=null, figureFileBig=null, tableContent=
毒力种类毒力因子基因名称(%)
细胞黏附Ⅲ型菌毛(Mrk)mrkA (97.37)、mrkB (97.37)、mrkC (97.37)、mrkD (97.37)、mrkF (97.37)、mrkH (97.37)、mrkI (94.74)、mrkJ (97.37)
Ⅰ型菌毛(Fim)fimA (100)、fimB (94.74)、fimC (100)、fimD (100)、fimE (100)、fimF (100)、fimG (100)、fimH (100)、fimI (100)、fimK (92.11)
外排泵ArcABarcA (100)、arcB (100)
铁摄取系统气杆菌素(Aer)iucA (42.11)、iucB (42.11)、iucC (42.11)、iucD (42.11)、iutA (100)
肠杆菌素(Ent)entA (100)、entB (100)、entC (100)、entD (100)、entE (100)、entF (100)、entS (100)、fepA (100)、fepB (100)、fepC (100)、fepD (100)、fepG (100)、fes (100)
沙门菌素(Sal)iroB (7.89)、iroC (5.26)、iroD (7.89)、iroE (100)、iroN (94.74)
耶尔森杆菌素(Ybt)fyuA (10.53)、irp1 (10.53)、irp2 (10.53)、ybtA (10.53)、ybtE (10.53)、ybtP (10.53)、ybtQ (10.53)、ybtS (10.53)、ybtT (10.53)、ybtU (10.53)、ybtX (10.53)
营养因子Allantoin utilizationallA (2.63)、allB (5.26)、allC (5.26)、allD (5.26)、allR (2.63)、allS (5.26)
黏液表型调控基因RcsAB、RmpArcsA (97.37)、rcsB (100)、rmpA (2.63)
毒素ColibactinclbA (2.63)、clbB (2.63)、clbC (0)、clbD (0)、clbE (2.63)、clbF (2.63)、clbG (2.63)、clbH (2.63)、clbI (2.63)、clbJ (2.63)、clbK (2.63)、clbL (2.63)、clbM (2.63)、clbN (2.63)、clbO (2.63)、clbP (2.63)、clbQ(2.63)、clbS (0)
), ArticleFig(id=1324078318479344355, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1324078309109269161, language=EN, label=Table 2, caption=

Antibiotic Resistance gene detection results

, figureFileSmall=null, figureFileBig=null, tableContent=
抗生素种类耐药基因(菌株数)株数携带率
(%)
β-内酰胺酶类blaSHV-108(1)、blaSHV-75(1)、blaSHV-42(1)、blaSHV-38(1)、blaSHV-27(3)、blaSHV-26(2)、blaSHV-14(1)、blaSHV-11(14)、blaSHV-1(9)、blaTEM-1(2)、blaOXA-10(5)、blaDHA-1(1)、blaOKP-A-12(1)、blaOKP-B-41(1)、blaLEN-2(1)、blaLEN-26(1)、blaLAP-2(11)38100.00
磷霉素类FosA6(28)、FosA5(7)3592.11
氨基糖苷类APH(6)-Id(14)、APH(3’’)-Ib(7)、AAC(3)-IId(1)、aadA(4)、aadA2(1)、CrcB(3)1842.36
四环素类tet(A)(15)1539.47
磺胺类sul1(1)、sul2(13)1334.21
喹诺酮类QnrS1(11)、QnrB4(1)1132.35
甲氧嘧啶类dfrA14(4)、dfrA12(1)、dfrA5(1)410.53
利福霉素类arr-2(4)410.53
酰胺醇类cmlA5(4)410.53
), ArticleFig(id=1324078318567424740, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1324078309109269161, language=CN, label=表2, caption=

耐药基因检出结果

, figureFileSmall=null, figureFileBig=null, tableContent=
抗生素种类耐药基因(菌株数)株数携带率
(%)
β-内酰胺酶类blaSHV-108(1)、blaSHV-75(1)、blaSHV-42(1)、blaSHV-38(1)、blaSHV-27(3)、blaSHV-26(2)、blaSHV-14(1)、blaSHV-11(14)、blaSHV-1(9)、blaTEM-1(2)、blaOXA-10(5)、blaDHA-1(1)、blaOKP-A-12(1)、blaOKP-B-41(1)、blaLEN-2(1)、blaLEN-26(1)、blaLAP-2(11)38100.00
磷霉素类FosA6(28)、FosA5(7)3592.11
氨基糖苷类APH(6)-Id(14)、APH(3’’)-Ib(7)、AAC(3)-IId(1)、aadA(4)、aadA2(1)、CrcB(3)1842.36
四环素类tet(A)(15)1539.47
磺胺类sul1(1)、sul2(13)1334.21
喹诺酮类QnrS1(11)、QnrB4(1)1132.35
甲氧嘧啶类dfrA14(4)、dfrA12(1)、dfrA5(1)410.53
利福霉素类arr-2(4)410.53
酰胺醇类cmlA5(4)410.53
), ArticleFig(id=1324078318672282341, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1324078309109269161, language=EN, label=Table 3, caption=

Antibiotic Resistance Status(n=38)

, figureFileSmall=null, figureFileBig=null, tableContent=
抗生素类别抗生素名称菌株数(耐药率,%)
合计预包装冷藏即食食品生猪肝
青霉素类氨苄西林31(81.58)19(86.36)12(75.0)
氨基糖苷类庆大霉素3(7.89)1(4.55)2(12.5)
阿米卡星---
四环素类四环素19(50.0)6(27.27)13(81.25)
替加环素---
苯丙醇类氯霉素17(44.74)4(18.18)13(81.25)
氟苯尼考17(44.74)4(18.18)13(81.25)
叶酸途径抑制剂磺胺甲噁唑/甲氧苄啶11(28.95)5(22.73)6(37.5)
β-内酰胺类
抗生素与β-内酰胺酶抑制剂氨苄西林/舒巴坦7(18.42)4(18.18)3(18.75)
头孢他啶/克拉维酸---
头孢噻肟/克拉维酸---
阿莫西林/克拉维酸1(2.63)-1(6.25)
头孢他啶/阿维巴坦---
头孢类头孢唑林4(10.53)2(9.09)2(12.5)
头孢西丁3(7.89)2(9.09)1(6.25)
头孢呋辛3(7.89)2(9.09)1(6.25)
头孢他啶---
头孢噻肟---
孢噻呋---
头孢吡肟---
喹诺酮类环丙沙星2(5.26)1(4.55)1(6.25)
萘啶酸---
乳菌肽脂肽类多粘菌素E1(2.63)-1(6.25)
多粘菌素B--
碳青霉烯类亚胺培南---
美罗培南---
厄他培南---
大环内酯类阿奇霉素---
), ArticleFig(id=1324078318772945638, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1324078309109269161, language=CN, label=表3, caption=

抗生素耐药情况(n=38)

, figureFileSmall=null, figureFileBig=null, tableContent=
抗生素类别抗生素名称菌株数(耐药率,%)
合计预包装冷藏即食食品生猪肝
青霉素类氨苄西林31(81.58)19(86.36)12(75.0)
氨基糖苷类庆大霉素3(7.89)1(4.55)2(12.5)
阿米卡星---
四环素类四环素19(50.0)6(27.27)13(81.25)
替加环素---
苯丙醇类氯霉素17(44.74)4(18.18)13(81.25)
氟苯尼考17(44.74)4(18.18)13(81.25)
叶酸途径抑制剂磺胺甲噁唑/甲氧苄啶11(28.95)5(22.73)6(37.5)
β-内酰胺类
抗生素与β-内酰胺酶抑制剂氨苄西林/舒巴坦7(18.42)4(18.18)3(18.75)
头孢他啶/克拉维酸---
头孢噻肟/克拉维酸---
阿莫西林/克拉维酸1(2.63)-1(6.25)
头孢他啶/阿维巴坦---
头孢类头孢唑林4(10.53)2(9.09)2(12.5)
头孢西丁3(7.89)2(9.09)1(6.25)
头孢呋辛3(7.89)2(9.09)1(6.25)
头孢他啶---
头孢噻肟---
孢噻呋---
头孢吡肟---
喹诺酮类环丙沙星2(5.26)1(4.55)1(6.25)
萘啶酸---
乳菌肽脂肽类多粘菌素E1(2.63)-1(6.25)
多粘菌素B--
碳青霉烯类亚胺培南---
美罗培南---
厄他培南---
大环内酯类阿奇霉素---
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贵州省食品中肺炎克雷伯菌毒力与耐药特征研究
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罗信旭 , 向婧姝 , 张德著 , 周倩 , 郑勤妮 , 周藜
现代预防医学 | 营养与食品卫生 2026,53(2): 252-258
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现代预防医学 |营养与食品卫生 2026 , 53 (2) : 252 -258
贵州省食品中肺炎克雷伯菌毒力与耐药特征研究
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罗信旭, 向婧姝, 张德著, 周倩, 郑勤妮, 周藜
作者信息
  • 贵州省疾病预防控制中心,贵州 贵阳 550004
通讯作者:
周藜,E-mail:
作者简介:

罗信旭(1990—),女,硕士,主管技师,研究方向:微生物学

Study on the virulence and drug-resistant characteristics of Klebsiella pneumoniae in foods sold,Guizhou
Xin-xu LUO, Jing-zhu XIANG, De-zhu ZHANG, Qian ZHOU, Qin-ni ZHENG, Li ZHOU
Affiliations
  • Guizhou provincial Center for Disease Control and Prevention, Guiyang, Guizhou 550004, China
出版时间: 2026-01-25 doi: 10.20043/j.cnki.MPM.202508323
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目的

了解贵州省市售食品中肺炎克雷伯菌分子分型、毒力与耐药特征,为我省有针对性的开展食品安全风险评估和监管决策提供科学依据。

方法

2023—2024年贵州省9个市(州)从市售预包装冷藏即食食品、生猪肝中分离的38株肺炎克雷伯菌进行拉丝试验、全基因组测序以及抗生素敏感性试验。

结果

38株分离株共筛选出4株高黏液型肺炎克雷伯菌(hypermucoviscous Klebsiella pneumoniae,hmKp)。通过全基因组序列分析,共获得20种荚膜型(capsule type,KL)、30种序列型(sequence typing,ST)、80种毒力基因和35种耐药基因。ST 37为主要菌株,分子型别为ST65-KL2的hmKp同时携带iucA、iroB、ybtS、rmpA、clb基因,ST490-KL3 肺炎克雷伯菌同时携带iucA、iroB、ybtS、allS基因。β-内酰胺酶类耐药基因以blaSHV-11、blaSHV-1、blaLAP-2为主,5株携带blaOXA-10基因。抗生素敏感性试验结果显示,耐药前五位依次是氨苄西林、四环素、氯霉素、氟苯尼考、磺胺甲噁唑/甲氧苄啶,耐药率分别为81.58%、50.0%、44.74%、44.74%和28.95%,对碳青霉烯类药物均敏感。

结论

贵州省市售食品中肺炎克雷伯菌毒力基因携带率高,致病力强,ST型和KL型呈现多样性。

食品  /  肺炎克雷伯菌  /  毒力基因  /  耐药
Objective

To investigate the molecular typing, virulence and antimicrobial resistance characteristics of food-borne Klebsiella pneumoniae in Guizhou Province, providing scientific evidence for targeted food safety risk assessment and regulatory decision-making in our province.

Methods

A total of 38 strains of Klebsiella pneumoniae were isolated from pre-packaged chilled ready-to-eat food and pig livers in 9 cities (states) of Guizhou Province from 2023 to 2024. Strains were screened by string test, whole genome sequencing and antimicrobial susceptibility test.

Results

Four hypermucoviscous Klebsiella pneumoniae strains were isolated from 38 isolates. Through whole genome sequence analysis, 20 KL types, 30 ST types, 80 virulence genes and 35 drug resistance genes were obtained. ST 37 was the main strain. HmKp of ST65-KL2 carried iucA, iroB, ybtS, rmpA and clb genes, while ST490-KL3 carried iucA, iroB, ybtS and allS genes. The main β-lactamase resistance genes were blaSHV-11, blaSHV-1, blaLAP-2. 5 of the strains carried blaOXA-10 gene. The results of antibiotic susceptibility tests showed that the top five resistant antibiotics were ampicillin, tetracycline, chloramphenicol, florfenicol and sulfamethoxazole/trimethoprim, with resistance rates of 81.58%, 50.0%, 44.74%, 44.74% and 28.95% respectively. All isolates were sensitive to carbapenems.

Conclusion

The prevalence of virulence genes in Klebsiella pneumoniae in food in Guizhou Province is high, with a strong pathogenic potencial. The ST types and KL types of the strains are diverse.

Food  /  Klebsiella pneumoniae  /  Virulence genes  /  Drug-resistant
罗信旭, 向婧姝, 张德著, 周倩, 郑勤妮, 周藜. 贵州省食品中肺炎克雷伯菌毒力与耐药特征研究. 现代预防医学, 2026 , 53 (2) : 252 -258 . DOI: 10.20043/j.cnki.MPM.202508323
Xin-xu LUO, Jing-zhu XIANG, De-zhu ZHANG, Qian ZHOU, Qin-ni ZHENG, Li ZHOU. Study on the virulence and drug-resistant characteristics of Klebsiella pneumoniae in foods sold,Guizhou[J]. Modern Preventive Medicine, 2026 , 53 (2) : 252 -258 . DOI: 10.20043/j.cnki.MPM.202508323
肺炎克雷伯菌(Klebsiella pneumoniae,Kp)是一种常见的革兰氏阴性条件致病菌,能够引起肺炎、肠炎、脑膜炎、败血症及爆发性紫癜[1]等严重疾病,是一种重要的人兽共患病原菌。有关数据显示,Kp已成为临床感染病原菌中仅次于大肠埃希菌的第二大致病菌。Kp感染治疗多选用 β -内酰胺类药物进行治疗,如头孢类和碳青霉烯类[2]。随着抗菌药物的广泛使用以及Kp具有多质粒和可移动遗传元件特性,病原菌耐药性不断增加。据CHINETS(https://www.chinets.com/)数据显示,从2005到2023年,Kp对亚胺培南、美罗培南的耐药率分别从3.0%、2.9%上升至24.8%、26.0%,耐药率显著增加。当今,肺炎克雷伯菌高致病性、高耐药性以及耐药菌株的有效治疗已成为全球关注的焦点。我国早在1996年就有Kp污染肉肠的报道[3],由Kp引起的食物中毒或暴发性事件也时有发生[4-5]。近年来,国内外学者不断从食品中分离出高毒力、高耐药性肺炎克雷伯菌[6-12],给食品带来了巨大安全隐患,给人类健康造成重大威胁。
预包装冷藏即食食品是一类可即食或经过简单加热后即可食用的产品,其味美、食用方便备受消费者喜爱。然而该类食品在制作和销售过程中由于环境、加工、贮藏及销售等因素的影响,极易受食源性致病菌污染,是导致我国食源性疾病暴发的主要食品种类之一。猪肝具有补血补铁作用,常食猪肝可防治眼睛干涩、疲劳,可调节和改善贫血病人的造血功能,还能帮助去除机体中的一些有毒成分。其加工方式多为爆炒,食材加工不彻底,存在一定的食用风险。课题组前期调查研究发现生猪肝中存在多种食源性致病菌,Hartantyo SHP.发现生猪肝中Kp检出率高达27%[13]。故本研究以市售预包装冷藏即食食品、生猪肝中分离的Kp作为研究对象,研究其毒力和耐药特征,旨在掌握我省食源肺炎克雷伯菌毒力及耐药现状,揭示其在食品安全中的风险隐患,为有针对性的开展风险评估和监管决策提供科学依据。
收集2023—2024年我省9个市(州)从市售预包装冷藏即食食品及生猪肝中分离的38株肺炎克雷伯菌,22株来源于预包装冷藏即食食品,16株为生猪肝中分离。所有菌株经分离纯培养后,挑取单菌落采用MALDI TOF MS质谱仪进行鉴定。
全自动微生物生化鉴别及药敏分析系统(VITEK 2 Compact,法国生物梅里埃公司)、基质辅助激光解吸电离飞行时间质谱(MALDI TOF MS,德国布鲁克公司)、革兰氏阴性菌药敏板(赛默飞世尔科技有限公司)。
分离株经过质谱仪鉴定后,挑取单个菌落划线接种至血琼脂平板,36 ℃培养24小时,用接种环向上挑取单个菌落。若挑起的黏液丝长度大于5mm,则判定为试验阳性,即为高黏液型肺炎克雷伯菌(hypermucoviscous Klebsiella pneumoniae,hmKp)。
采用革兰阴性菌药敏板及微量肉汤稀释法对分离株进行最小抑菌浓度(minimum inhibitory concentration,MIC)测定。药敏板包含11大类28种抗菌药物,药敏结果依据美国临床和实验室标准协会(clinical and laboratory standard institute,CLSI)要求进行折点判断。同时设立阴性和空白对照,以大肠埃希氏菌ATCC 25922作为质控。
使用细菌基因组DNA提取试剂盒(MagPure Bacterial DNA Kit)提取核酸,1%琼脂糖凝胶电泳确认DNA完整性,合格的核酸送至生工生物工程(上海)股份有限公司进行高通量测序。文库构建采用DNA建库试剂盒(Hieff NGS® MaxUp Ⅱ DNA Library Prep Kit for Illumina®),华大智造DNBseq-T7测序仪进行二代测序,读长为PE150。使用Trimmomatic(v0.36)对测序数据进行质控,BV-BRC在线网站(https://www.bv-brc.org/)进行序列拼接组装。
分别使用PubMLST网站、毒力因子数据库(virulence factors database,VFDB)、综合性抗生素耐药性数据库(comprehensive antibiotic resistance database,CARD)在线分析菌株序列分型(sequence typing,ST)、荚膜型(capsule type,KL)、wzi基因、毒力基因和耐药基因。使用CSI Phylogeny 1.4(https://cge.food.dtu.dk/services/CSIPhylogeny/)绘制菌株单核苷酸多态性(single nucleotide polymorphism,SNP)系统发育树。使用Chiplot网站(https://www.chiplot.online/)对系统发育树进行可视化和美化。
数据的统计分析采用SPSS 19.0软件,率的比较采用χ2检验,P<0.05为差异有统计学意义。
通过拉丝试验共检出4株hmKp菌株,阳性率为10.51%,菌株全部来源于市售预包装冷藏即食食品。
通过PubMLST数据库在线比对,38株Kp共发现30种ST型,两株新ST型。其中ST37型别4株,均来自猪肝,ST17和ST34各2株,ST31、ST36、ST45、ST46、ST65、ST133、ST186、ST281、ST416、ST447、ST461、ST490、ST515、ST540、ST610、ST753、ST899、ST950、ST1155、ST1446、ST1727、ST1876、ST1916、ST2402、ST3393、ST4110、ST5388各1株。通过wzi基因在线比对分析共发现20种KL型,其中从市售预包装冷藏即食食品中检出一株高致病血清型KL2毒力株。4株hmKp分子型别组合为ST65-KL2、ST1155-KL25、ST416-KL20、ST2402-KL22/37/8/30/151,详见图1。
利用VFDB数据库对38株菌Kp进行毒力基因预测,共获得80种毒力相关基因,分别与细胞黏附(Mrk、Fim)、外排泵(AcrAB)、铁摄取系统(Ent、Sal、Ybt、Aer)、黏液表型调控(RcsAB、RmpA)、营养因子(All)和外毒素Colibactin相关,详见表1。所有菌株均携带fimA、fimH、arcA、arcB、entB、fepA基因,检出率为100%。mrkA、mrkD、iucA、fyuA、ybtS、iroB、allS、rmpA、clb基因检出率分别为97.37%、97.37%、42.11%、10.53%、10.53%、7.89%、5.26%、2.63%、2.63%。其中猪肝iucA基因检出率(14/16,87.5%)明显高于预包装冷藏即食食品(2/22,9.09%),差异有统计学意义(χ2=23.362,P<0.001)。其中1株分子型别组合为ST65-KL2 的hmKp同时携带iucA、iroB、ybtS、rmpA、clb基因,分子型别组合为ST490-KL3同时携带iucA、iroB、ybtS、allS基因,详见图2。
通过综合抗生素耐药性数据库(CARD)进行耐药基因在线分析显示,38株Kp共检出9大类35种耐药基因,详见表2。其中β-内酰胺酶类耐药基因17种,占比43.59%,以blaSHV-11、blaLAP-2、blaSHV-1、blaOXA-10为主,检出率分别为36.84%、28.95%、23.68%、13.16%。所有菌株至少携带1种β-内酰胺酶类耐药基因,其中1株菌同时携带5种β-内酰胺酶类耐药基因。磷霉素类以FosA6为主,氨基糖苷类以APH(6)-Id(14/38)为主,四环素类只检出tet(A)基因,喹诺酮类以QnrS1(11/38)为主,磺胺类以sul2为主。
采用微量肉汤稀释法对38株分离株进行抗菌药物敏感性试验,结果显示耐药率前五依次为氨苄西林、四环素、氯霉素、氟苯尼考、磺胺甲噁唑/甲氧苄啶,耐药率分别为81.58%、50.0%、44.74%、44.74%和28.95%,对碳青霉烯类药物均敏感。其中生猪肝中肺炎克雷伯菌对四环素、氯霉素、氟苯尼考耐药率显著高于预包装冷藏即食食品,差异有统计学意义(其中四环素耐药率:χ2=10.795,P=0.003;氯霉素、氟苯尼考耐药率:χ2=14.903,P<0.001),结果详见表3。多重耐药菌株占比达42.11%,其中1株生猪肝来源的KP对12种抗生素同时耐药,详见图3。
肺炎克雷伯菌可分为经典肺炎克雷伯菌(classic Klebsiella pneumoniae,cKp)和高毒力肺炎克雷伯菌(hypervirulent Klebsiella pneumoniae,hvKp)常见于社区和医院获得性感染,是肠杆菌科克雷伯菌属中一类重要的条件致病菌。MLST分子分型被证实是判断hvKp毒力水平的良好指标[14]。亚洲地区Kp以ST11为主[15],ST11同时也是我国临床分离株的优势菌株[16],而我省临床分离株主要为ST23[17]。目前食品中已报道主要有ST101[10]、ST1049[18]、ST76[19]等。本研究ST37为主要型别,均来源于生猪肝,该研究结果与盛席静对猪源Kp的研究结果一致[20]。ST37广范存在于全球人源和动物源中,具有一定的传播风险。hvKp常伴随高粘液表型,过去研究者常选用拉丝试验来判断菌株是否属于hvKp。随着研究的深入,发现部分hvKp并不具有高黏液表型,因此不能仅凭高黏液表型来区分cKp和hvKp。肺炎克雷伯菌的毒力与荚膜多糖(Capsule Polysaccharides, CPS)、脂多糖(Lipopolysaccharide, LPS)、菌毛及铁载体密切相关[21]。CPS由cps基因簇(如galF、wzi、wza)编码,其中,wzi基因编码的外膜蛋白wzi参与荚膜多糖附着于外膜,是重要的毒力因子之一。不同荚膜多糖变体称为K抗原,其中Kl、K2、K5、K20、K54、K57被认为是hvKp的主要血清型。本研究从预包装冷藏即食食品中发现一株KL2型高黏液型肺炎克雷伯菌,分子型别组合为ST65-KL2,该型别组合的Kp常以高毒力的形式出现在临床分离株中[22]。Kp高黏液表型通常认为主要与rmpA基因有关,但本研究高黏液型Kp中rmpA基因携带率仅为25%,说明rmpA基因并不是高黏液表型的决定性因素,该结果与严伟等人研究结果一致[23]。Kp可分泌多种铁载体,研究发现90%以上hvKp均检出Ybt、Aer和Sal。Ybt的分泌和转运主要由ybtS和fyuA基因编码,Aer和Sal分别由iucABCD-intA和iroBCDE-iroN基因簇编码,其中iucA和iroB常作为hvKp的生物标记之一。本研究iucA、iroB、ybtS和fyuA基因检出率分别为42.11%、7.89%、10.53%、10.53%,3株同时携带iucA和iroB基因,2株同时携带iucA、iroB、ybtS和fyuA,猪肝中iucA基因检出率显著高于预包装冷藏即食食品,提示生猪肝中肺炎克雷伯菌的毒力更高。2株携带allS基因,该基因可使hvKp在营养缺乏的条件下以尿素作为碳、氮源,从而提高hvKp生长定植能力。值得注意的是,分子型别为ST65-KL2的hmKp同时携带iucA、iroB、ybtS、fyuA、rmpA、clb基因。clb基因簇可编码colibactin基因毒素,从而导致细菌感染的真核细胞发生DNA双链损伤诱发癌变[24]。
随着抗生素的广泛使用,细菌的耐药性不断增强。研究显示2016~2020年我国临床高毒力耐碳青霉烯类肺炎克雷伯菌检出率从28.2%增长至45.7%[25]。产超广谱β-内酰胺酶(Extended Spectrum Beta-Lactamases,ESBLs)和耐碳青霉烯类(Carbapenem-Resistant,CR)肺炎克雷伯菌已被WHO列为全球公共卫生的最大威胁之一。肺炎克雷伯菌对β -内酰胺类抗生素产生耐药性的主要原因是可产生β -内酰胺酶。β -内酰胺酶主要包括ESBLs、头孢菌素酶和碳青霉烯酶,其中常见的ESBLs耐药基因有blaSHV、blaTEM、blaCTX-M,碳青霉烯酶耐药基因有blaKPC、blaVIM、blaNDM、blaIMP、blaOXA。这些耐药基因常位于可移动元件内,可在人类、动物及环境之间进行传播。本研究发现所有菌株均携带β -内酰胺酶类药物耐药基因,其中blaSHV基因携带率最高,高达87.2%。shv型β -内酰胺酶耐药基因普遍存在于Kp,导致该菌对多种β -内酰胺类抗生素天然耐药,特别是氨苄西林。此外,研究还发现了blaOXA-10型碳青霉烯酶耐药基因。目前食品中Kp分离株常携带的碳青霉烯酶耐药基因主要有blaOXA-48、blaIMP 、blaNDM、blaKPC、blaVIM[26-27],以上碳青霉烯酶耐药基因同时也是临床上常见的。
抗生素敏感性试验发现分离株对9大类13种抗生素产生不同层次耐药,对碳青霉烯酶类、大环内酯类高度敏感。前五位依次为氨苄西林、四环素、氯霉素、氟苯尼考、磺胺甲噁唑/甲氧苄啶,耐药率分别为81.58%、50.0%、44.74%、44.74%和28.95%。庆大霉素、氨苄西林/舒巴坦、阿莫西林/克拉维酸、头孢唑林、头孢西丁、头孢呋辛、环丙沙星、多粘菌素E表现处不同层次的耐药。该研究结果与国外研究结果有所不同,Ashwath P.[28]发现新鲜蔬菜和鸡肉中的Kp分离株对头孢噻肟、四环素完全耐药,但对氯霉素、磺胺甲噁唑/甲氧苄啶保持高度敏感。Bedair HM.[29]发现蔬菜沙拉中Kp分离株对头孢呋辛、头孢拉定完全耐药,对四环素、氯霉素、磺胺甲噁唑/甲氧苄啶耐药率均低于26%。本研究Kp分离株对四环素、氯霉素、氟苯尼考、磺胺甲噁唑/甲氧苄啶的耐药性高于我国生牛乳分离株[18-19],可见不同食品来源、不同地区的菌株耐药性存在一定差异。Kot B.[30]等人发现家禽来源Kp对青霉素类、III和IV代头孢菌素类耐药率较高,而猪源分离株对第一、二代头孢菌素、大环内酯类抗生素表现出高耐药性,对四环素类耐药率高于50%。Li Y[21]等人研究发现我国西北地区Kp对头孢唑林、氯霉素和磺胺甲噁唑的耐药率低于北部和南部地区。生猪肝中Kp对四环素、氯霉素、氟苯尼考耐药率显著高于预包装冷藏即食食品,提示在家畜养殖过程中可能存在抗生素的不合理使用。该研究结果对家畜Kp感染的临床用药具有一定指导意义。此外,本次研究中多重耐药菌株占比高达42.11%,其中猪肝多重耐药率达68.75%,提示畜肉可能是多重耐药菌的储存库。本研究部分抗生素耐药基因与耐药表型呈现较好的吻合性,如氨苄西林、四环素、磺胺甲噁唑/甲氧苄啶,而氯霉素、氟苯尼考的耐药基因携带率远低于耐药率,说明苯丙醇类抗生素的耐药除耐药基因的介导外,还存在其他耐药途径。
综上所述,贵州省市售预包装冷藏即食食品和生猪肝中肺炎克雷伯菌表现出一定的毒力和耐药性。鉴于研究试验菌株相对较少,菌株时间、空间分布较窄以及食物的加工方式,理应加强对此类食品的监测,以便及时发现风险隐患,从一定程度上保障食品安全。
  • 贵州省科技支撑计划项目(黔科合支撑[2021]一般435号)
  • 贵州省卫生健康委科学技术基金项目(gzwkj2025-513)
  • 贵州省微生物组与传染性疾病防控重点实验室(ZDSYS【2023】004)
  • 贵州省传染病预防与控制人才基地科研团队(RCJD2105)
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doi: 10.20043/j.cnki.MPM.202508323
  • 接收时间:2025-08-22
  • 首发时间:2026-11-02
  • 出版时间:2026-01-25
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  • 收稿日期:2025-08-22
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贵州省科技支撑计划项目(黔科合支撑[2021]一般435号)
贵州省卫生健康委科学技术基金项目(gzwkj2025-513)
贵州省微生物组与传染性疾病防控重点实验室(ZDSYS【2023】004)
贵州省传染病预防与控制人才基地科研团队(RCJD2105)
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    贵州省疾病预防控制中心,贵州 贵阳 550004

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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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