临床来源无乳链球菌血清型分布和基因组特征研究

Serotype distribution and genomic characteristics of clinical Streptococcus agalactiae strains

  • 摘要:
    目的 了解临床来源无乳链球菌的血清型分布及基因组特征,为防治无乳链球菌感染提供依据。
    方法 对本实验室2016—2023年收集的177株临床来源无乳链球菌,通过多重聚合酶链式反应(PCR)方法进行血清型鉴定,采用PCR方法进行序列型别(ST)分型,并对每种血清型中随机选取的26株菌进行高通量基因组测序。 基因组数据用于血清型分型验证、ST分型、系统发育分析及毒力和耐药基因预测。
    结果 PCR分型鉴定出6种血清型(Ⅰa、Ⅰb、Ⅱ、Ⅲ、Ⅳ和Ⅴ),Ⅰb、Ⅴ和Ⅲ型占90.40%;26株菌基因组分型共鉴定出7种血清型,其中1株菌未能分型,部分菌株分型结果与PCR分型结果不一致。 177株菌株中检出23种ST、8个克隆复合群;进行系统发育分析发现Ⅰb型菌株具有更高的遗传保守性;共预测出54个毒力基因,涉及黏附、外毒素等功能;耐药基因共16个,其中mreASaga_mprF为构成菌株基础耐药机制的基因,较多菌株携带四环素耐药基因tetM和大环内酯类耐药基因ermB
    结论 本研究明确了无乳链球菌的血清型、ST分布及基因组特征,并揭示了其毒力与耐药基因分布特征。 未来需加强对无乳链球菌血清型分布及耐药性的监测,优化分型方法,为疫苗开发及感染防控策略提供科学依据。

     

    Abstract:
    Objective To understand the serotype distribution and genomic characteristics of clinical Streptococcus agalactiae strains, and provide evidence for the prevention and treatment of S. agalactiae infection.
    Methods A total of 177 clinical strains of S. agalactiae, collected in our laboratory between 2016 and 2023, were used for serotype identification with multiplex polymerase chain reaction (PCR), sequence type (ST) identification was conducted using PCR, and 26 strains were randomly selected from each serotype for high-throughput genome sequencing. The genomic data were used for serotype verification, ST typing, phylogenetic analysis, and the prediction of virulence and resistance genes.
    Results PCR identified six serotypes (Ⅰa, Ⅰb, Ⅱ, Ⅲ, Ⅳ, and Ⅴ), with Ⅰb, Ⅴ, and Ⅲ types accounting for 90.40%. Genomic typing of 26 strains revealed 7 serotypes (1 untyped), some strains had inconsistent typing results with PCR. In the 177 strains, 23 STs and 8 clonal complexes were detected. The 26 sequenced strains were classified into 13 distinct STs. Phylogenetic analysis showed that serotype Ⅰb strains exhibited greater genetic conservation. A total of 54 virulence genes were predicted, which are associated with functions such as adhesion and exotoxin production. The strains carried 16 antimicrobial resistance genes, with mreA and Saga_mprF constituting core genetic determinants of basal resistance. Many strains carried tetracycline resistance gene tetM and macrolide resistance gene ermB.
    Conclusion This study analyzed the predominant serotypes (types Ⅰb, Ⅴ, and Ⅲ), STs and genomic characteristics of S. agalactiae, and revealed the distribution patterns of its virulence and resistance genes. It is essential to strengthen the surveillance for S. agalactiae serotype distribution and drug resistance, improve typing methodologies in the future to provide evidence for vaccine development and infection prevention strategies and reduce the disease burden associated with S. agalactiae infection.

     

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