基于宏基因组测序的孕晚期B族链球菌不同定植状况阴道微环境比较分析

Comparison of vaginal microbiota in pregnant women with different group B streptococcus colonization statuses in the third timester based on metagenomic sequencing

  • 摘要:
    目的 探究孕晚期B族链球菌(GBS)不同定植状况阴道微生物的物种组成、功能、耐药情况及差异,为GBS定植与阴道微生态之间的相互作用与影响提供基础数据。
    方法 收集孕晚期接受GBS产筛孕妇的阴道拭子样本。通过临床微生物常规培养筛选GBS培养阳性的样本,选取同期孕妇基本情况相似,但GBS培养阴性的样本作为对照,提取选取样本的DNA,完成宏基因组序列测定,采用生物信息学分析比较GBS阳性组和阴性组阴道微生物种群、代谢及耐药相关基因的组成与差异,探讨GBS定植对阴道菌群的影响。
    结果 在150份样品中共有13份样品GBS培养阳性,选取相匹配的16份GBS培养阴性样本作为对照,宏基因组测序分析显示:GBS阳性组阴道微生物多样性显著低于阴性组(P<0.05)。微生物差异分析表明,GBS阴性组以卷曲乳杆菌(LDA=5.274, P<0.01)和詹氏乳杆菌(LDA=4.072, P<0.05)等保护性乳杆菌为主,而GBS阳性组则显著富集惰性乳杆菌(LDA=5.528, P<0.001),且条件致病菌比例升高。两组在微生物功能与代谢通路上存在显著差异(P<0.05),在大环内酯类、多肽类及大环内酯-林可酰胺-链阳菌素B(MLS)类等抗菌药物耐药基因类别上也存在显著差异(P<0.05),其中大环内酯类耐药基因ermB在GBS阳性组中的相对丰度显著高于阴性组(P<0.05)。
    结论 孕晚期不同GBS定植状况的阴道微环境有明显差异,阳性定植的阴道微环境种群多样性降低,有益乳杆菌丰度减少,耐药基因的相对丰度增加,提示孕期GBS定植可能与阴道微环境相关,通过改善阴道微环境,有可能减少GBS的阴道定植,保障母婴健康。

     

    Abstract:
    Objective To investigate the species composition, functionality, and drug resistance of vaginal microbiota under different Group B Streptococcus (GBS) colonization statuses in late pregnancy, and to provide foundational data for understanding the interaction and impact between GBS colonization and the vaginal microecology.
    Methods Vaginal swab samples were collected from pregnant women undergoing prenatal GBS screening in late pregnancy. Samples positive for GBS were identified through conventional clinical microbiological culture. Simultaneously, samples negative for GBS culture from pregnant women with similar baseline characteristics were selected as controls. DNA was extracted from the selected samples, and metagenomic sequencing was performed. Bioinformatics analysis was used to compare the composition and differences in vaginal microbial populations, metabolism, and resistance-related genes between the GBS-positive and GBS-negative groups, aiming to explore the impact of GBS colonization on the vaginal microbiota.
    Results Among 150 samples, 13 tested positive for GBS culture. Matched 16 GBS culture-negative samples were selected as controls. Metagenomic sequencing analysis revealed that the diversity of vaginal microbiota in the GBS-positive group was significantly lower than that in the negative group (P<0.05). Microbial differential analysis indicated that the GBS-negative group was predominantly colonized by protective Lactobacillus species such as Lactobacillus crispatus (LDA = 5.274, P<0.01) and Lactobacillus jensenii (LDA= 4.072, P<0.05). In contrast, the GBS-positive group showed significant enrichment of Lactobacillus iners (LDA=5.528, P<0.001), with an increased proportion of opportunistic pathogens. Significant differences were observed between the two groups in microbial functions and metabolic pathways (P<0.05), as well as in categories of antibiotic resistance genes, including macrolides, polypeptides, and Macrolide-Lincosamide-Streptogramin B (MLS) (P<0.05). Specifically, the relative abundance of the macrolide resistance gene ermB was significantly higher in the GBS-positive group compared to the negative group (P<0.05).
    Conclusions The vaginal microenvironment in late pregnancy differs significantly based on GBS colonization status. GBS-positive colonization is associated with reduced microbial diversity, decreased abundance of beneficial Lactobacillus species, and increased relative abundance of resistance genes. These findings suggest that GBS colonization during pregnancy may be related to the vaginal microenvironment. Improving the vaginal microenvironment could potentially reduce vaginal GBS colonization, thereby safeguarding maternal and neonatal health.

     

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