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Article(id=1320438068318945530, tenantId=1146029695717560320, journalId=1319373219989389464, issueId=1320438006813672182, articleNumber=null, orderNo=null, doi=10.11847/zgggws1146556, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1733155200000, receivedDateStr=2024-12-03, revisedDate=1757865600000, revisedDateStr=2025-09-15, acceptedDate=1760371200000, acceptedDateStr=2025-10-14, onlineDate=1792746568039, onlineDateStr=2026-10-23, pubDate=1773072000000, pubDateStr=2026-03-10, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1792746568039, onlineIssueDateStr=2026-10-23, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1792746568039, creator=13701087609, updateTime=1792746568039, updator=13701087609, issue=Issue{id=1320438006813672182, tenantId=1146029695717560320, journalId=1319373219989389464, year='2026', volume='42', issue='3', pageStart='257', pageEnd='384', issueExtLink='null', onlineDate='null', pubDate='1773072000000', pubDateStr='2026-03-10', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1792746553376, creator='13701087609', updateTime=1792748100383, updator='13701087609', preIssue=null, nextIssue=null, articleTotal=null, ext={EN=IssueExt(id=1320444495519404406, tenantId=1146029695717560320, journalId=1319373219989389464, issueId=1320438006813672182, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1320444495519404407, tenantId=1146029695717560320, journalId=1319373219989389464, issueId=1320438006813672182, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null, downloadFileDto=null}, startPage=348, endPage=353, ext={EN=ArticleExt(id=1320438068549632251, articleId=1320438068318945530, tenantId=1146029695717560320, journalId=1319373219989389464, language=EN, title=Molecular biological characteristics of Streptococcus pneumoniae serotype 15B isolated from children in Zaozhuang city: a study based on surveillance data in sentinel hospitals from 2022 to 2023, columnId=1320427686514770171, journalTitle=Chinese Journal of Public Health, columnName=Survey and Report, runingTitle=null, highlight=null, articleAbstract=
Objective

To understand the prevalence of Streptococcus pneumoniae (SP) serotypes among children in Zaozhuang, Shandong from 2022 to 2023 and study the antibiotic resistance mechanism, pathogenic mechanis, and MLST typing characteristics of SP serotype 15B strains by whole genome sequencing, thus providing data support for vaccine development and clinical antibiotic treatment.

Methods

A total of 87 SP isolates from children (≤ 14 years old) infected with pneumonia in Zaozhuang from 2022 to 2023 were collected. PCR was used for serotype testing. Antimicrobial sensitivity tests, whole genome sequencing, and bioinformatics analysis were conducted for SP serotype 15B strains.

Results

The SP serotypes prevalent in this region were mainly 19F (41.4%), 6B (18.4%), 15B (13.8%), and 23F (11.5%). The serotype coverage rate of the 13-valent pneumococcal conjugate vaccine (PCV13) was 77%, and that of 23-valent pneumococcal polysaccharide vaccine (PPSV23) was 92%. SP serotype 15B had a resistance rate of 100% to clindamycin, erythromycin, tetracycline, and compound sulfamethoxazole, with the multi-drug resistance rate of 100%. SP serotype 15B carried resistance genes ermB, rlmA(Ⅱ), tetM, patA, patB, and pmrA. The resistance genes (ermB, rlmA(Ⅱ), and tetM) and resistance phenotypes were consistent. All the SP serotype 15B strains carried virulence genes cbpG, pce/cbpE, pavA, pfbA, lmb, slrA, plr/gapA, cbpD, lytA, lytB, lytC, hysA, nanA, nanB, eno, cps4A, cps4B, cps4C, cps4D, zmpB, iga, cppA, htrA/degP, tig/ropA, piaA, psaA, piuA, and ply. Most (83.3%) of them carried pspA, and a small portion (16.7%) carried pspC/cbpA, srtB, srtC, srtD, rrgA, rrgB, and rrgC. ST9785 was the main sequence type of SP serotype 15B in this region.

Conclusions

The serotype 15B is the predominant PPSV23 serotype among the strains not covered by PCV13, and ST9785 is the main sequence type. SP serotype 15B in this region is fully resistant to clindamycin, erythromycin, tetracycline, and methoxybenzylamine/sulfamethoxazole and shows severe multi-drug resistance.

, authors=Huihui LI, Yi ZHAO, Lili WANG, Xueru YIN, Pin LIU, authorsList=Huihui LI, Yi ZHAO, Lili WANG, Xueru YIN, Pin LIU, authorCompany=null, correspAuthors=Lili WANG, Pin LIU, authorNote=null, correspAuthorsNote=
WANG Lili，E-mail:
LIU Pin, E-mail:
, copyrightStatement=Copyright © 2026 Chinese Journal of Public Health. All rights reserved., 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=1320438069149417726, articleId=1320438068318945530, tenantId=1146029695717560320, journalId=1319373219989389464, language=CN, title=枣庄地区儿童感染15B型肺炎链球菌分子生物学特征分析：基于2022—2023年哨点医院监测数据, columnId=1320427686770622717, journalTitle=中国公共卫生, columnName=调查报告与分析, runingTitle=null, highlight=null, articleAbstract=
目的

了解2022—2023年山东省枣庄地区儿童感染肺炎链球菌（SP）血清型流行情况，利用全基因组测序技术分析15B型SP的耐药机制、致病机理和多位点序列分型（MLST）特点，为疫苗研发和临床抗生素治疗提供数据支持。

方法

收集枣庄地区2022—2023年≤14岁儿童感染肺炎患者的87株SP分离株，使用聚合酶链式反应（PCR）技术进行SP血清型检测，对15B型SP进行药敏试验、全基因组测序和生物信息学分析。

结果

枣庄地区流行的SP血清型主要是19 F（41.4%）、6 B（18.4%）、15 B（13.8%）和23 F（11.5%），13价肺炎球菌结合疫苗（PCV13）疫苗血清型覆盖率为77%，23价肺炎球菌多糖疫苗（PPSV23）疫苗血清型覆盖率为92%；15B型SP对克林霉素、红霉素、四环素和复方新诺明耐药率均为100%，多重耐药率为100%，携带耐药基因ermB、rlmA（Ⅱ）、tetM、patA、patB和pmrA，其中耐药基因ermB、rlmA（Ⅱ）和tetM与耐药表型一致；15B型SP全部携带毒力基因cbpG、pce/cbpE、pavA、pfbA、lmb、slrA、plr/gapA、cbpD、lytA、lytB、lytC、hysA、nanA、nanB、eno、cps4A、cps4B、cps4C、cps4D、zmpB、iga、cppA、htrA/degP、tig/ropA、piaA、psaA、piuA和 ply；大部分（83.3%）携带pspA基因；小部分（16.7%）携带pspC/cbpA、srtB、srtC、srtD、rrgA、rrgB、rrgC。ST 9785是本地区15B型SP的主要型别。

结论

15B型是枣庄地区非PCV 13血清型覆盖的菌株血清型中主要流行的PPSV 23血清型，主要基因型为ST 9785，15B型SP对克林霉素、红霉素、四环素和复方新诺明全部耐药，并出现严重的多重耐药现象。

, authors=李会会, 赵毅, 王莉莉*, *, 尹雪茹, 刘品*, *, authorsList=李会会, 赵毅, 王莉莉, 尹雪茹, 刘品, authorCompany=null, correspAuthors=王莉莉, 刘品, authorNote=

李会会（1986—），副主任技师，硕士，研究方向：病原微生物检测

, correspAuthorsNote=
王莉莉，E-mail：
刘品，E-mail：
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李会会（1986—），副主任技师，硕士，研究方向：病原微生物检测

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李会会（1986—），副主任技师，硕士，研究方向：病原微生物检测

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Structural and functional insights into peptidoglycan access for the lytic amidase LytA of Streptococcus pneumoniae[J]. mBio, 2014, 5(1): e01120-e01213., articleTitle=null, refAbstract=null)], funds=null, companyList=[AuthorCompany(id=1320438069388493055, tenantId=1146029695717560320, journalId=1319373219989389464, articleId=1320438068318945530, xref=null, ext=[AuthorCompanyExt(id=1320438069396881664, tenantId=1146029695717560320, journalId=1319373219989389464, articleId=1320438068318945530, companyId=1320438069388493055, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=Department of Pathogenic Microbiology, Zaozhuang Center for Disease Control and Prevention, Zaozhuang 277000, China), AuthorCompanyExt(id=1320438069409464577, tenantId=1146029695717560320, journalId=1319373219989389464, articleId=1320438068318945530, companyId=1320438069388493055, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=枣庄市疾病预防控制中心病原微生物科，枣庄 277000)])], figs=[ArticleFig(id=1320438072009933097, tenantId=1146029695717560320, journalId=1319373219989389464, articleId=1320438068318945530, language=EN, label=Fig.1, caption=Cluster analysis of SP serotype 15B strains based on MLST results, figureFileSmall=FgjOURhsOeMkM8oWzzeepg==, figureFileBig=kUDAo+vc3iLVnNBMdLS5vg==, tableContent=null), ArticleFig(id=1320438072102207786, tenantId=1146029695717560320, journalId=1319373219989389464, articleId=1320438068318945530, language=CN, label=图1, caption=15B型SP MLST聚类分析结果, figureFileSmall=FgjOURhsOeMkM8oWzzeepg==, figureFileBig=kUDAo+vc3iLVnNBMdLS5vg==, tableContent=null), ArticleFig(id=1320438072316117291, tenantId=1146029695717560320, journalId=1319373219989389464, articleId=1320438068318945530, language=EN, label=Tab.1, caption=

Resistance and resistance genes of SP serotype 15B to 14 antibiotics

, figureFileSmall=null, figureFileBig=null, tableContent=
菌株名称抗生素耐药情况耐药基因比对一致性/%
AMXFEPCTXCHLCMELVXLZDMEMMXFPTETSXTVAermBrlmA（Ⅱ）patApatBpmrAtetM
ZZ0848SPSSSSRRSSSSSRRS99.87100100100100100
ZZ0459SPRIISRRSSRSIRRS99.87100100100100100
ZZ0460SPIISSRRSSISSRRS99.87100100100100100
ZZ0465SPIISSRRSSRSIRRS99.87100100100100100
ZZ0472SPIISSRRSSRSIRRS99.87100100100100100
ZZ0814SPRISSRRSSRSIRRS99.87100100100100100
ZZ0639SPRIISRRSSRSIRRS99.87100100100100100
ZZ0074SPSSISRRSSISRRRS99.87100100100100100
ZZ0149SPSSSSRRSSISRRRS99.87100100100100100
ZZ0156SPSSISRRSSISRRRS99.87100100100100100
ZZ0627SPRIISRRSSRSRRRS99.87100100100100100
ZZ0637SPRRISRRSSRSRRRS99.87100100100100100
耐药率/%41.78.30.00.0100.0100.00.00.058.30.041.7100.0100.00.0
), ArticleFig(id=1320438072408391980, tenantId=1146029695717560320, journalId=1319373219989389464, articleId=1320438068318945530, language=CN, label=表1, caption=

15 B SP对14种抗生素的耐药性和耐药基因携带情况

, figureFileSmall=null, figureFileBig=null, tableContent=
菌株名称抗生素耐药情况耐药基因比对一致性/%
AMXFEPCTXCHLCMELVXLZDMEMMXFPTETSXTVAermBrlmA（Ⅱ）patApatBpmrAtetM
ZZ0848SPSSSSRRSSSSSRRS99.87100100100100100
ZZ0459SPRIISRRSSRSIRRS99.87100100100100100
ZZ0460SPIISSRRSSISSRRS99.87100100100100100
ZZ0465SPIISSRRSSRSIRRS99.87100100100100100
ZZ0472SPIISSRRSSRSIRRS99.87100100100100100
ZZ0814SPRISSRRSSRSIRRS99.87100100100100100
ZZ0639SPRIISRRSSRSIRRS99.87100100100100100
ZZ0074SPSSISRRSSISRRRS99.87100100100100100
ZZ0149SPSSSSRRSSISRRRS99.87100100100100100
ZZ0156SPSSISRRSSISRRRS99.87100100100100100
ZZ0627SPRIISRRSSRSRRRS99.87100100100100100
ZZ0637SPRRISRRSSRSRRRS99.87100100100100100
耐药率/%41.78.30.00.0100.0100.00.00.058.30.041.7100.0100.00.0
), ArticleFig(id=1320438072479695149, tenantId=1146029695717560320, journalId=1319373219989389464, articleId=1320438068318945530, language=EN, label=Tab.2, caption=

Virulence genes carried by SP serotype 15B

, figureFileSmall=null, figureFileBig=null, tableContent=
毒力基因
类别		主要毒力
基因		毒力基因比对一致性/%携带率/%
ZZ0848SPZZ0074SPZZ0149SPZZ0156SPZZ0459SPZZ0460SPZZ0465SPZZ0472SPZZ0627SPZZ0637SPZZ0639SPZZ0814SP
黏附相关cbpG100100100100100100100100100100100100100.0
pce/cbpE100100100100100100100100100100100100100.0
pspC/cbpA—————100———100——16.7
pavA100100100100100100100100100100100100100.0
pfbA100100100100100100100100100100100100100.0
srtB100——————————10016.7
srtC100——————————10016.7
srtD100——————————10016.7
rrgA100——————————10016.7
rrgB100——————————10016.7
rrgC100——————————10016.7
lmb100100100100100100100100100100100100100.0
slrA100100100100100100100100100100100100100.0
plr/gapA100100100100100100100100100100100100100.0
胞外酶cbpD100100100100100100100100100100100100100.0
lytA100100100100100100100100100100100100100.0
lytB100100100100100100100100100100100100100.0
lytC100100100100100100100100100100100100100.0
hysA100100100100100100100100100100100100100.0
nanA100100100100100100100100100100100100100.0
nanB100100100100100100100100100100100100100.0
eno100100100100100100100100100100100100100.0
免疫调节cps4A100100100100100100100100100100100100100.0
cps4B100100100100100100100100100100100100100.0
cps4C100100100100100100100100100100100100100.0
cps4D100100100100100100100100100100100100100.0
pspA—100100100100—10010010010010010083.3
zmpB100100100100100100100100100100100100100.0
iga100100100100100100100100100100100100100.0
cppA100100100100100100100100100100100100100.0
htrA/degP100100100100100100100100100100100100100.0
tig/ropA100100100100100100100100100100100100100.0
营养代谢因子溶血素相关psaA100100100100100100100100100100100100100.0
piaA100100100100100100100100100100100100100.0
piuA100100100100100100100100100100100100100.0
ply100100100100100100100100100100100100100.0
), ArticleFig(id=1320438072571969838, tenantId=1146029695717560320, journalId=1319373219989389464, articleId=1320438068318945530, language=CN, label=表2, caption=

15B型SP的毒力基因携带情况

, figureFileSmall=null, figureFileBig=null, tableContent=
毒力基因
类别		主要毒力
基因		毒力基因比对一致性/%携带率/%
ZZ0848SPZZ0074SPZZ0149SPZZ0156SPZZ0459SPZZ0460SPZZ0465SPZZ0472SPZZ0627SPZZ0637SPZZ0639SPZZ0814SP
黏附相关cbpG100100100100100100100100100100100100100.0
pce/cbpE100100100100100100100100100100100100100.0
pspC/cbpA—————100———100——16.7
pavA100100100100100100100100100100100100100.0
pfbA100100100100100100100100100100100100100.0
srtB100——————————10016.7
srtC100——————————10016.7
srtD100——————————10016.7
rrgA100——————————10016.7
rrgB100——————————10016.7
rrgC100——————————10016.7
lmb100100100100100100100100100100100100100.0
slrA100100100100100100100100100100100100100.0
plr/gapA100100100100100100100100100100100100100.0
胞外酶cbpD100100100100100100100100100100100100100.0
lytA100100100100100100100100100100100100100.0
lytB100100100100100100100100100100100100100.0
lytC100100100100100100100100100100100100100.0
hysA100100100100100100100100100100100100100.0
nanA100100100100100100100100100100100100100.0
nanB100100100100100100100100100100100100100.0
eno100100100100100100100100100100100100100.0
免疫调节cps4A100100100100100100100100100100100100100.0
cps4B100100100100100100100100100100100100100.0
cps4C100100100100100100100100100100100100100.0
cps4D100100100100100100100100100100100100100.0
pspA—100100100100—10010010010010010083.3
zmpB100100100100100100100100100100100100100.0
iga100100100100100100100100100100100100100.0
cppA100100100100100100100100100100100100100.0
htrA/degP100100100100100100100100100100100100100.0
tig/ropA100100100100100100100100100100100100100.0
营养代谢因子溶血素相关psaA100100100100100100100100100100100100100.0
piaA100100100100100100100100100100100100100.0
piuA100100100100100100100100100100100100100.0
ply100100100100100100100100100100100100100.0
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枣庄地区儿童感染15B型肺炎链球菌分子生物学特征分析：基于2022—2023年哨点医院监测数据
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李会会 , 赵毅 , 王莉莉 *, * , 尹雪茹 , 刘品 *, *
中国公共卫生 | 调查报告与分析 2026,42(3): 348-353
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中国公共卫生 |调查报告与分析 2026 , 42 (3) : 348 -353
枣庄地区儿童感染15B型肺炎链球菌分子生物学特征分析：基于2022—2023年哨点医院监测数据
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李会会, 赵毅, 王莉莉*, * , 尹雪茹, 刘品*, *
作者信息
  • 枣庄市疾病预防控制中心病原微生物科，枣庄 277000
通讯作者:
王莉莉，E-mail：
刘品，E-mail：
作者简介:

李会会（1986—），副主任技师，硕士，研究方向：病原微生物检测

Molecular biological characteristics of Streptococcus pneumoniae serotype 15B isolated from children in Zaozhuang city: a study based on surveillance data in sentinel hospitals from 2022 to 2023
Huihui LI, Yi ZHAO, Lili WANG , Xueru YIN, Pin LIU
Affiliations
  • Department of Pathogenic Microbiology, Zaozhuang Center for Disease Control and Prevention, Zaozhuang 277000, China
    • Corresponding Author:
    WANG Lili，E-mail:
    LIU Pin, E-mail:
出版时间: 2026-03-10 doi: 10.11847/zgggws1146556
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摘要
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目的

了解2022—2023年山东省枣庄地区儿童感染肺炎链球菌（SP）血清型流行情况，利用全基因组测序技术分析15B型SP的耐药机制、致病机理和多位点序列分型（MLST）特点，为疫苗研发和临床抗生素治疗提供数据支持。

方法

收集枣庄地区2022—2023年≤14岁儿童感染肺炎患者的87株SP分离株，使用聚合酶链式反应（PCR）技术进行SP血清型检测，对15B型SP进行药敏试验、全基因组测序和生物信息学分析。

结果

枣庄地区流行的SP血清型主要是19 F（41.4%）、6 B（18.4%）、15 B（13.8%）和23 F（11.5%），13价肺炎球菌结合疫苗（PCV13）疫苗血清型覆盖率为77%，23价肺炎球菌多糖疫苗（PPSV23）疫苗血清型覆盖率为92%；15B型SP对克林霉素、红霉素、四环素和复方新诺明耐药率均为100%，多重耐药率为100%，携带耐药基因ermB、rlmA（Ⅱ）、tetM、patA、patB和pmrA，其中耐药基因ermB、rlmA（Ⅱ）和tetM与耐药表型一致；15B型SP全部携带毒力基因cbpG、pce/cbpE、pavA、pfbA、lmb、slrA、plr/gapA、cbpD、lytA、lytB、lytC、hysA、nanA、nanB、eno、cps4A、cps4B、cps4C、cps4D、zmpB、iga、cppA、htrA/degP、tig/ropA、piaA、psaA、piuA和 ply；大部分（83.3%）携带pspA基因；小部分（16.7%）携带pspC/cbpA、srtB、srtC、srtD、rrgA、rrgB、rrgC。ST 9785是本地区15B型SP的主要型别。

结论

15B型是枣庄地区非PCV 13血清型覆盖的菌株血清型中主要流行的PPSV 23血清型，主要基因型为ST 9785，15B型SP对克林霉素、红霉素、四环素和复方新诺明全部耐药，并出现严重的多重耐药现象。

15B型肺炎链球菌  /  血清型  /  耐药性  /  耐药基因  /  毒力基因  /  多位点序列分型（MLST）  /  枣庄市
Objective

To understand the prevalence of Streptococcus pneumoniae (SP) serotypes among children in Zaozhuang, Shandong from 2022 to 2023 and study the antibiotic resistance mechanism, pathogenic mechanis, and MLST typing characteristics of SP serotype 15B strains by whole genome sequencing, thus providing data support for vaccine development and clinical antibiotic treatment.

Methods

A total of 87 SP isolates from children (≤ 14 years old) infected with pneumonia in Zaozhuang from 2022 to 2023 were collected. PCR was used for serotype testing. Antimicrobial sensitivity tests, whole genome sequencing, and bioinformatics analysis were conducted for SP serotype 15B strains.

Results

The SP serotypes prevalent in this region were mainly 19F (41.4%), 6B (18.4%), 15B (13.8%), and 23F (11.5%). The serotype coverage rate of the 13-valent pneumococcal conjugate vaccine (PCV13) was 77%, and that of 23-valent pneumococcal polysaccharide vaccine (PPSV23) was 92%. SP serotype 15B had a resistance rate of 100% to clindamycin, erythromycin, tetracycline, and compound sulfamethoxazole, with the multi-drug resistance rate of 100%. SP serotype 15B carried resistance genes ermB, rlmA(Ⅱ), tetM, patA, patB, and pmrA. The resistance genes (ermB, rlmA(Ⅱ), and tetM) and resistance phenotypes were consistent. All the SP serotype 15B strains carried virulence genes cbpG, pce/cbpE, pavA, pfbA, lmb, slrA, plr/gapA, cbpD, lytA, lytB, lytC, hysA, nanA, nanB, eno, cps4A, cps4B, cps4C, cps4D, zmpB, iga, cppA, htrA/degP, tig/ropA, piaA, psaA, piuA, and ply. Most (83.3%) of them carried pspA, and a small portion (16.7%) carried pspC/cbpA, srtB, srtC, srtD, rrgA, rrgB, and rrgC. ST9785 was the main sequence type of SP serotype 15B in this region.

Conclusions

The serotype 15B is the predominant PPSV23 serotype among the strains not covered by PCV13, and ST9785 is the main sequence type. SP serotype 15B in this region is fully resistant to clindamycin, erythromycin, tetracycline, and methoxybenzylamine/sulfamethoxazole and shows severe multi-drug resistance.

Streptococcus pneumoniae serotype 15B  /  serotype  /  antibiotic resistance  /  antibiotic resistance genes  /  virulence genes  /  multi-locus sequence typing (MLST)  /  Zaozhuang city
李会会, 赵毅, 王莉莉, 尹雪茹, 刘品. 枣庄地区儿童感染15B型肺炎链球菌分子生物学特征分析：基于2022—2023年哨点医院监测数据. 中国公共卫生, 2026 , 42 (3) : 348 -353 . DOI: 10.11847/zgggws1146556
Huihui LI, Yi ZHAO, Lili WANG, Xueru YIN, Pin LIU. Molecular biological characteristics of Streptococcus pneumoniae serotype 15B isolated from children in Zaozhuang city: a study based on surveillance data in sentinel hospitals from 2022 to 2023[J]. Chinese Journal of Public Health, 2026 , 42 (3) : 348 -353 . DOI: 10.11847/zgggws1146556
正文
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肺炎链球菌（Streptococcus pneumoniae, SP）是一种革兰氏阳性球菌，是全球细菌性肺炎和脑膜炎的主要病因，可导致菌血症、急性中耳炎、鼻窦炎、气管炎等疾病[1]，被认为是儿童呼吸道细菌感染最重要的致病菌之一。在<5岁儿童中，SP是致死率较高病原体[2]。迄今为止，已经发现46个血清群中的100多个血清型[3]。接种肺炎链球菌结合疫苗能够显著降低肺炎链球菌疾病的发病率，但是也导致致病性SP血清型分布的变化[4–6]，因此持续监测SP血清型的分布变化和病原特征具有重要意义。目前国内对15B型SP的研究较少，而山东枣庄地区更是缺乏对15B型SP的研究。本研究通过监测山东省枣庄地区2022—2023年SP分离株的血清型分布，利用全基因组测序技术对15B型SP进行耐药机制、致病机理和MLST序列特征分析，有助于预防和控制SP感染，为疫苗研发和临床治疗提供数据支持。
1　材料与方法
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1.1　菌种来源
样本来自山东枣庄市2家三级甲等医院（枣庄市妇幼保健院和滕州市中心人民医院）2022年6月—2023年11月因感染肺炎就诊的儿童患者，本研究涉及的临床样本相关信息已通过枣庄市疾病预防控制中心伦理委员会的审核（批准号：ZZCDCLL2025002），菌株编号与临床数据关联信息仅限研究组成员在保密条件下访问。病例纳入标准：（1）确诊为肺炎，肺炎病例应具备急性感染表现；（2）年龄≤14岁；（3）从临床样本中分离和鉴定出SP 菌株；（4）未接种肺炎链球菌相关疫苗。排除同一患者多次入院和连续分离自同一患者的菌株。将收集的SP疑似菌株接种于血平板，在5% CO2，37 ℃的培养箱培养18～20 h，利用微生物质谱仪对培养物进行复核确证，最终获得87株SP分离株。试验中使用的质控菌株大肠埃希菌 ATCC25922和肺炎链球菌ATCC49619均购自美国菌种保藏中心。
1.2　主要仪器及试剂
血平板（北京陆桥技术股份有限公司）；肺炎链球菌核酸多重PCR血清分型检测试剂盒（北京卓诚惠生生物科技股份有限公司）；链球菌药敏检测板（美国Thermo公司）；FastPure Bacteria DNA Isolation Mini Kit基因组提取试剂盒（南京诺唯赞生物科技股份有限公司）；QuantStudio5荧光定量PCR仪、全自动微生物鉴定和药敏分析系统和CO2培养箱（美国ThermoFisher公司）；毛细管电泳仪（德国QIAGEN公司）；微生物质谱仪MBT Compass（德国BRUKER公司），以上试剂均经过验收合格后在有效期内使用，仪器经过检定校准合格后投入使用。
1.3　实验方法
1.3.1　PCR分型鉴定
SP分离株的纯培养物提取DNA，利用PCR技术，通过A、B、C、D 4个PCR反应进行核酸扩增，用毛细管电泳仪对核酸扩增产物进行电泳分析，根据扩增条带的大小对常见的23种血清型（血清型1、2、3、4、5、6 B、7 F、8、9 N、9 V、10 A、11 A、12 F、14、15 B、17 F、18 C、19 A、19 F、20、22 F、23 F和33 F）SP进行分型。
1.3.2　药敏试验
采用微量肉汤法进行药物敏感性试验，根据链球菌药敏检测板使用说明书并结合菌落特性，将菌悬液调至0.5麦氏浊度，接种至药敏板后，使用全自动微生物鉴定和药敏分析仪进行培养和结果判读。使用大肠埃希氏菌ATCC25922进行质控。根据美国临床实验室标准化协会（Clinical and Laboratory Standards Institute, CLSI）标准[7]判定SP对以下14种抗生素的敏感（S）、中度敏感（I）和耐药（R）结果：阿莫西林（amoxicillin, AMX）、头孢吡肟（cefepime, FEP）、头孢噻肟（cefotaxime, CTX）、氯霉素（chloramphenicol, CHL）、克林霉素（clindamycin, CM）、红霉素（erythromycin, E）、左氧氟沙星（levofloxacin, LVX）、利奈唑胺（linazolamide, LZD）、美罗培南（meropenem, MEM）、莫西沙星（moxifloxacin, MXF）、青霉素（penicillin, P）、四环素（tetracycline, TET）、复方新诺明（methoxybenzylamine/sulfamethoxazole, SXT）、万古霉素（vancomycin, VA）。
1.3.3　全基因组测序
采用FastPure Bacteria DNA Isolation Mini Kit基因组提取试剂盒提取SP分离株基因组DNA，使用NovaSeq X Plus平台进行SP地建库测序，并对原始数据进行去除接头和低质量reads处理以及对测序数据进行质控（由北京诺禾致源科技股份有限公司完成），使用CLC Genomics Workbench 22.0.2分析软件进行测序数据拼接工作，获得SP分离株的基因组序列。
1.3.4　耐药基因及毒力基因分析
将15B型SP分离株基因组拼接序列分别上传耐药基因数据库（The Comprehensive Antibiotic Resistance Database, CARD）（https://card.mcmaster.ca/）和毒力因子数据库（Virluence Factor Database, VFDB）（http://www.mgc.ac.cn/VFs/），在线进行相关耐药基因和毒力基因的预测。
1.3.5　MLST序列特征分析及聚类分析
将15B型SP分离株基因组拼接序列上传到PubMLST分子分型数据库中的Streptococcus pneumoniae 模块平台（https://pubmlst.org/organisms/streptococcus-pneumoniae），进行相应的管家基因的比对，获得菌株的MLST分型，利用MEGA 5.2.2进行系统发育树构建。
2　结　果
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2.1　SP血清型别分布情况
2022年6月—2023年11月，从感染肺炎儿童患者痰液样本中共分离获得87株SP分离株，进行多重PCR分析，有80株（92%）可以鉴定出血清型，7株（8%）SP分离株的血清型不在试剂盒检测范围内，未做分型。分析结果显示19F（41.4%）是主要血清型，其次为6 B（18.4%）、15 B（13.8%）和23 F（11.5%），包含少量的18 C（2.3%）、22 F（1.1%）、7 F（1.1%）、14（1.1%）型和3（1.1%）型血清型。PCV13疫苗血清型覆盖率为77%，PPSV 23疫苗血清型覆盖率为92%，在非PCV 13血清型覆盖的菌株中，主要流行的PPSV23血清型为15 B。
2.2　全基因组测序结果
12株15B型SP经全基因组测序组装完成的细菌基因组框架图碱基大小为2.07～2.13 Mbp，contig数为22～41个，G+C含量为39.51%～39.62%。
2.3　15B型SP的耐药情况（表1）
药敏试验显示，12株15B型SP对克林霉素、红霉素、四环素和复方新诺明耐药率均为100%。对头孢噻肟、氯霉素、左氧氟沙星、利奈唑胺、莫西沙星和万古霉素全部敏感。阿莫西林和青霉素的耐药率为41.7%（5/12），美罗培南耐药率58.3%（7/12），头孢吡肟耐药率8.3%（1/12），多重耐药率均为100%。使用耐药基因数据库CARD对以上菌株的全基因序列进行耐药基因分析，发现12株15B型SP的耐药基因携带情况高度一致，均携耐药基因ermB、rlmA（Ⅱ）和tetM，耐药基因与耐药表型一致。
2.4　15B型SP的毒力基因携带情况（表2）
使用的毒力因子数据库主要包括胞外酶基因、溶血素基因、免疫调节基因、营养代谢因子、黏附相关基因等。经统计分析，12株15B型SP都携带毒力基因cbpG、pce/cbpE、pavA、pfbA、lmb、slrA、plr/gapA、cbpD、lytA、lytB、lytC、hysA、nanA、nanB、eno、cps4A、cps4B、cps4C、cps4D、zmpB、iga、cppA、htrA/degP、tig/ropA、piuA、psaA、piaA和ply基因。83.3%（10/12）菌株携带pspA基因；仅16.7%（2/12）菌株携带pspC/cbpA、srtB、srtC、srtD、rrgA、rrgB、rrgC。
2.5　15B型SP的MLST序列特征分析及聚类分析（图1）
根据PubMLST网站，对7个管家基因在线分析，12株15B型SP的MLST分型分别为ST9785、ST3397、ST6555和ST10088。利用MEGA软件构建系统发育树，可见12株15B型SP聚类为4个cluster，最主要的cluster为ST9785型，是本地区15B型SP主要ST型，占比75%（9/12）。
3　讨　论
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本研究显示，山东省枣庄市儿童感染SP主要是19 F型，占到全部菌株的41.4%，其次为6 B型、15B型和23 F型，PCV13疫苗的血清型覆盖率为77%， PPSV23疫苗的血清型覆盖率高达92%。有研究报告显示，自从发展中国家开始接种PCV13以来，15 B 型SP的分离率一直在增加，在中国儿童中，非PCV13涵盖的15B型SP很常见，并且15 B是主要血清型，应当引起重视[8]。在本地区流行的血清型中，15B型检出率高达13.8%，是枣庄地区非PCV 13血清型覆盖的菌株血清型中，主要流行的PPSV23血清型。与柳州市的研究结果一致[9]，因此应该加强SP的血清型监测，持续关注15群SP的流行状况，并优化肺炎链球菌疫苗的血清型组成，提高其覆盖度。
枣庄地区的15B型SP对克林霉素、红霉素、四环素和复方新诺明耐药率100%，并对4～6种抗生素同时耐药，以克林霉素（林可酰胺类）+红霉素（大环内酯类）+四环素+复方新诺明（磺胺类）的耐药模式为主，与苏州市和柳州市的研究结果一致[9–10]，多重耐药现象严重，为临床治疗带来困难。从分子水平了解SP的耐药机制，对解决其耐药性问题至关重要，深入研究其耐药情况和机制可为临床用药提供数据支持。通过分析其全基因组序列的耐药基因，发现该15B型SP均携带耐药基因ermB、rlmA（Ⅱ）、tetM、patA、patB和pmrA。其中ermB是红霉素耐药相关基因，由其介导的核糖体甲基化修饰是SP对大环内酯类耐药的最主要机制[11]；tetM是四环素类抗生素耐药相关基因，可阻止四环素与核糖体结合而失去抑菌作用，导致四环素出现耐药[12]；rlmA（Ⅱ）存在于许多革兰氏阳性细菌中，可使23S rRNA甲基化，对大环内酯类和林可酰胺类抗生素产生耐药性[13]。ermB、rlmA（Ⅱ）、tetM耐药基因与耐药表型一致率均为100%。本研究中15B型SP全部携带耐药基因patA、patB和pmrA，却均对氟喹诺酮类抗生素（左氧氟沙星和莫西沙星）敏感，耐药基因与耐药表型高度不一致。这是由于虽然耐药基因patA、patB和pmrA可以介导药物外排，引起喹诺酮类药物耐药的产生，但是喹诺酮类药物耐药机制与抗菌靶点突变有关，喹诺酮类耐药决定区的基因突变才是产生耐药性的主要原因[14]。
SP致病能力和毒力因子密切相关，SP常见的毒力因子包括荚膜多糖和黏附因子，如胞外酶、IgA1蛋白酶、金属离子吸收因子、蛋白酶和毒素因子，这些因子参与了 SP相关疾病的发病机制[15]。深入研究SP的毒力基因不仅可以了解其致病力还可以为开发下一代蛋白质疫苗提供关键候选靶点，极高的携带率表明了开发疫苗的潜力[16]。本地区的15B型SP主要分别携带了29～35个毒力基因，其中胞外酶基因（cbpD、lytA、lytB、lytC、hysA、nanA、nanB、eno）、溶血素基因（ply）、免疫调节基因（cps4A、cps4B、cps4C、cps4D、zmpB、iga、cppA、htrA/degP、tig/ropA）、营养代谢因子（psaA、piaA、piuA）、黏附相关基因（cbpG、pce/cbpE、pavA、pfbA、lmb、slrA、plr/gapA）的携带率为100%，pspA携带率83.3%，pspC/cbpA、rrgA、rrgB、rrgC、srtB、srtC和srtD携带率16.7%。pspA是SP的菌体表面蛋白，可结合人乳铁蛋白，具有抑制补体系统活化的功能，pspA的缺失将会导致SP定植、入侵能力的下降[17]。pspC/cbpA基因表达的肺炎链球菌表面蛋白C（PSPC）是菌株侵入肺部及血液系统的必要毒力因子[18]。rrgA、rrgB、rrgC、srtB、srtC和srtD属于rlrA基因毒力岛，主要调控肺炎链球菌PI–1菌毛结构的表达[9]。溶血素基因ply是SP的外毒素相关基因，其产物为肺炎球菌溶血素，导致炎症的发生[19]。lytA基因主要编码自溶酶A，可促进毒素和免疫逃避因子的传播，易被青霉素和万古霉素激活，介导细菌自溶从而导致毒素及其他毒力因子的释放，使病情加重[20]，因此治疗携带lytA基因的SP应避免使用青霉素和万古霉素抗生素。
对15B型SP的7个管家基因在线分析发现MLST分型分别为ST9785、ST3397、ST6555和ST10088，其中75%的15B型SP基因分型为ST9785，是本地区主要流行的15B型SP基因型。经数据分析发现本地区相同ST型的15B型SP，携带的毒力基因基本一致，耐药基因完全一致，和其他ST型略有差异。
本研究通过分析枣庄地区SP的血清型分布，了解当地SP流行情况和PCV13疫苗的实际血清型覆盖情况，发现15B型SP是本地区流行的非PCV13疫苗血清型，并出现了严重的多重耐药现象，应当持续监测。由于收集的SP分离株数量有限，通过有限的数据只能对当地SP流行状况和病原特征有初步的了解，后续还需要不断加强监测以建立本地区的SP全基因组数据库，为细菌性传染病的防控提供数据支持。

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doi: 10.11847/zgggws1146556
  • 接收时间：2024-12-03
  • 首发时间：2026-10-23
  • 出版时间：2026-03-10
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  • 收稿日期：2024-12-03
  • 修回日期：2025-09-15
  • 录用日期：2025-10-14
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    枣庄市疾病预防控制中心病原微生物科，枣庄 277000

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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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