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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

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

    摘要
  • 摘要:
    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。

     

    Abstract:
    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.

     

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