细菌性阴道病及其合并盆腔炎患者的阴道菌群分析
|
摘要
目的对比细菌性阴道病及其合并盆腔炎患者与健康个体的阴道菌群,分析阴道菌群的结构,为确定该疾病的特征细菌及研究致病机制奠定基础。方法采用临床Amsel标准筛选的17例细菌性阴道病患者、13例细菌性阴道病合并盆腔炎患者和52例健康者,使用无菌拭子采集阴道后穹窿分泌物以提取细菌基因组DNA。采用PCR技术扩增上一步得到的16S rRNA片段,而后将扩增产物通过变形梯度凝胶电泳(DGGE)分离以得到阴道细菌种属结构图谱,应用Quantity One软件进行聚类分析,应用凝胶测序法进行特异条带分析。检测16S rRNA基因,具体研究样品中的物种分类。结果细菌性阴道病及其合并盆腔炎患者的阴道菌群的构成与健康者相比较具有显著性差异。其中,Firmicutes等菌门细菌减少,Actinobacteria等菌门细菌增多;G.vaginallis、P.vaginallis等厌氧菌数量均增加,L.crispatus、L.iners等益生菌数量均有减少。结论细菌性阴道病及其合并盆腔炎患者的生殖道内的合并感染改变了阴道内的原有微生态平衡。两组疾病组患者的阴道菌群构成相比于健康对照组变化明显。
Objective To analyze the structure of vaginal microbiota of patients with bacterial vaginosis and those with concurrent pelvic inflammatory disease vs healthy individuals, and lay a foundation for identifying the characteristics of the disease-specific bacteria and studying the pathogenesis. Methods 17 patients with bacterial vaginosis, 13 patients with bacterial vaginosis complicated with pelvic inflammatory disease, and 52 healthy women were screened by using clinical Amsel criteria. Sterile swabs were used to collect vaginal secretions from the posterior fornix to extract bacterial genomic DNA. The 16 S rRNA fragment was amplified with PCR, then the amplified products were separated using deformation gradient gel electrophoresis(DGGE) to obtain the structure map of vaginal bacterial species. Cluster analysis was carried out with Quantity One software, and the specific bands were analyzed with gel sequencing. The 16 S rRNA gene was detected to specifically study the classification of species in the samples. Results Compared with the control group, the composition of vaginal microflora in patients with vaginal diseases changed significantly. Among them, the number of bacteria such as Firmicutes and Actinobacteria increased, the number of anaerobic bacteria such as G. vaginallis and P. vaginallis increased, while the number of probiotics such as L. crispatus and L. iners decreased. Conclusion Bacterial vaginosis and co-infection in genital tract of patients with pelvic inflammatory disease change the original microecological balance in vagina. The vaginal microbiota composition in disease groups was significantly different from that in healthy control group.
Objective To analyze the structure of vaginal microbiota of patients with bacterial vaginosis and those with concurrent pelvic inflammatory disease vs healthy individuals, and lay a foundation for identifying the characteristics of the disease-specific bacteria and studying the pathogenesis. Methods 17 patients with bacterial vaginosis, 13 patients with bacterial vaginosis complicated with pelvic inflammatory disease, and 52 healthy women were screened by using clinical Amsel criteria. Sterile swabs were used to collect vaginal secretions from the posterior fornix to extract bacterial genomic DNA. The 16 S rRNA fragment was amplified with PCR, then the amplified products were separated using deformation gradient gel electrophoresis(DGGE) to obtain the structure map of vaginal bacterial species. Cluster analysis was carried out with Quantity One software, and the specific bands were analyzed with gel sequencing. The 16 S rRNA gene was detected to specifically study the classification of species in the samples. Results Compared with the control group, the composition of vaginal microflora in patients with vaginal diseases changed significantly. Among them, the number of bacteria such as Firmicutes and Actinobacteria increased, the number of anaerobic bacteria such as G. vaginallis and P. vaginallis increased, while the number of probiotics such as L. crispatus and L. iners decreased. Conclusion Bacterial vaginosis and co-infection in genital tract of patients with pelvic inflammatory disease change the original microecological balance in vagina. The vaginal microbiota composition in disease groups was significantly different from that in healthy control group.
引文
[1] Verstraelen H,Verhelst R.Bacterial vaginosis:An update on diagnosis and treatment[J].Expert Rev Anti Infect Ther,2009,7(9):1109-1124.
[2] Lewis WG,Robinson LS,Perry J,et al.Hydrolysis of secreted sialoglycoprotein immunoglobulin A(IgA) in vivo and biochemical models of bacterial vaginosis[J].J Biol Chem,2012,287(3):2079-2089.
[3] 王丹丹,毕芳芳,杨清.腹腔镜在诊断和治疗盆腔炎症性疾病方面的应用[J].国际妇产科学杂志,2014,41(5):555-557.
[4] 施新野.妇科盆腔炎症90例临床分析[J].中外医学研究,2014,12(30):137-138.
[5] 李珂,李明传,尹伶,等.120例急性盆腔炎症性疾病患者的致病因素及微生物特点的临床分析[J].中国性科学,2018,27(2):57-60.
[6] Oh HY,Kim BS,Seo SS,et al.The association of uterine cervical microbiota with an increased risk for cervical intraepithelial neoplasia in Korea[J].Clin Microbiol Infect,2015,21(7):674.e1-9.
[7] Ravel J,Gajer P,Abdo Z,et al.Vaginal microbiome of reproductive-age women[J].Proc Natl Acad Sci U S A,2011,108(Suppl 1):4680-4687.
[8] Gajer P,Brotman RM,Bai G,et al.Temporal dynamics of the human vaginal microbiota[J].Sci Transl Med,2012,4(132):132ra52.
[9] O′hanlon DE,Moench TR,Cone RA.Vaginal pH and microbicidal lactic acid when lactobacilli dominate the microbiota[J].PLoS One,2013,8(11):e80074.
[10] Jones IS.A histological assessment of normal vulval skin[J].Clin Exp Dermatol,1983,8(5):513-521.
[11] Yeoman CJ,Thomas SM,Miller ME,et al.A multi-omic systems-based approach reveals metabolic markers of bacterial vaginosis and insight into the disease[J].PLoS One,2013,8(2):e56111.
[12] Doerflinger SY,Throop AL,Herbst-Kralovetz MM.Bacteria in the vaginal microbiome alter the innate immune response and barrier properties of the human vaginal epithelia in a species-specific manner[J].J Infect Dis,2014,209(12):1989-1999.
[13] Rose WA,Mcgowin CL,Spagnuolo RA,et al.Commensal bacteria modulate innate immune responses of vaginal epithelial cell multilayer cultures[J].PLoS One,2012,7(3):e32728.
[14] Van De Wijgert JH,Borgdorff H,Verhelst R,et al.The vaginal microbiota:What have we learned after a decade of molecular characterization?[J].PLoS One,2014,9(8):e105998.
[15] Huang B,Fettweis JM,Brooks JP,et al.The changing landscape of the vaginal microbiome[J].Clin Lab Med,2014,34(4):747-761.
[16] Gajer P,Brotman RM,Bai G,et al.Temporal dynamics of the human vaginal microbiota[J].Sci Transl Med,2012,4:132ra52-132ra52.
[2] Lewis WG,Robinson LS,Perry J,et al.Hydrolysis of secreted sialoglycoprotein immunoglobulin A(IgA) in vivo and biochemical models of bacterial vaginosis[J].J Biol Chem,2012,287(3):2079-2089.
[3] 王丹丹,毕芳芳,杨清.腹腔镜在诊断和治疗盆腔炎症性疾病方面的应用[J].国际妇产科学杂志,2014,41(5):555-557.
[4] 施新野.妇科盆腔炎症90例临床分析[J].中外医学研究,2014,12(30):137-138.
[5] 李珂,李明传,尹伶,等.120例急性盆腔炎症性疾病患者的致病因素及微生物特点的临床分析[J].中国性科学,2018,27(2):57-60.
[6] Oh HY,Kim BS,Seo SS,et al.The association of uterine cervical microbiota with an increased risk for cervical intraepithelial neoplasia in Korea[J].Clin Microbiol Infect,2015,21(7):674.e1-9.
[7] Ravel J,Gajer P,Abdo Z,et al.Vaginal microbiome of reproductive-age women[J].Proc Natl Acad Sci U S A,2011,108(Suppl 1):4680-4687.
[8] Gajer P,Brotman RM,Bai G,et al.Temporal dynamics of the human vaginal microbiota[J].Sci Transl Med,2012,4(132):132ra52.
[9] O′hanlon DE,Moench TR,Cone RA.Vaginal pH and microbicidal lactic acid when lactobacilli dominate the microbiota[J].PLoS One,2013,8(11):e80074.
[10] Jones IS.A histological assessment of normal vulval skin[J].Clin Exp Dermatol,1983,8(5):513-521.
[11] Yeoman CJ,Thomas SM,Miller ME,et al.A multi-omic systems-based approach reveals metabolic markers of bacterial vaginosis and insight into the disease[J].PLoS One,2013,8(2):e56111.
[12] Doerflinger SY,Throop AL,Herbst-Kralovetz MM.Bacteria in the vaginal microbiome alter the innate immune response and barrier properties of the human vaginal epithelia in a species-specific manner[J].J Infect Dis,2014,209(12):1989-1999.
[13] Rose WA,Mcgowin CL,Spagnuolo RA,et al.Commensal bacteria modulate innate immune responses of vaginal epithelial cell multilayer cultures[J].PLoS One,2012,7(3):e32728.
[14] Van De Wijgert JH,Borgdorff H,Verhelst R,et al.The vaginal microbiota:What have we learned after a decade of molecular characterization?[J].PLoS One,2014,9(8):e105998.
[15] Huang B,Fettweis JM,Brooks JP,et al.The changing landscape of the vaginal microbiome[J].Clin Lab Med,2014,34(4):747-761.
[16] Gajer P,Brotman RM,Bai G,et al.Temporal dynamics of the human vaginal microbiota[J].Sci Transl Med,2012,4:132ra52-132ra52.