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[Illumina测序16S rRNA标签揭示与细菌性阴道病相关的多样阴道微生物群]

[Illumina sequencing 16S rRNA tagging reveals diverse vaginal microbiomes associated with bacterial vaginosis].

作者信息

Xu Surong, Zong Lili, Liu Mubiao, He Yan, Huang Xuemei, Zhou Hongwei

机构信息

Department of Obstetrics and Gynecology, Southern Medical University, Guangzhou, China.

出版信息

Nan Fang Yi Ke Da Xue Xue Bao. 2013 May;33(5):672-7.

Abstract

OBJECTIVE

To analyze and compare vaginal microbiomes in healthy women at child-bearing ages and patients with bacterial vaginosis (BV).

METHODS

A total of 74 vaginal swabs of the vaginal fornix were collected from 37 BV patients and 37 healthy women. BV status was assessed according to Amsels clinical criteria for all the subjects and confirmed using Gram-stain criteria (Nugent scores). Genomic DNA of the samples was extracted for amplifying the 16S rRNA V6 hypervariable region by PCR and pyrosequencing by Illumina. BIPES, UCHIME, TSC and GAST were employed to analyze the information of the species from the samples.

RESULTS

Lactobacillus was the predominant species in healthy women (more than 95%), including mainly L. iners and L. crispatus, with a small quantity of Gardnerella, Granulicatella, Streptococcus, Prevotella, Escherichia and other genus. The α diversity was significantly increased in 30 BV patients (P<0.001), and β diversity also changed obviously shown by decreased Lactobacillus (varying from 45% to 1%, consisting mainly of L. iners) or even absence Lactobacillus in 6 cases, with increased relative abundance of Gardnerella, Prevotella, Granulicatella, Anaerococcus, Parvimonas, Peptoniphilus.harei, Peptostreptococcus, and Dialister. Different from previous data, 7 BV cases showed a predominance of the rare species L.gasseri and L.acidophilus (75% to 50%).

CONCLUSION

Lactobacillus is the predominant vaginal species in healthy women (mainly L. iners and L. crispatus) co-existing with many other bacteria and a variety of microorganisms. Lactobacillus is significantly decreased and even absent in most of BV patients, and some cases show the predominance of the rare species L.gasseri and L.acidophilus.

摘要

目的

分析和比较育龄期健康女性与细菌性阴道病(BV)患者的阴道微生物群。

方法

从37例BV患者和37例健康女性中收集了74份阴道穹窿部的阴道拭子。根据阿姆斯尔临床标准对所有受试者评估BV状态,并使用革兰氏染色标准(纽金特评分)进行确认。提取样本的基因组DNA,通过PCR扩增16S rRNA V6高变区,并采用Illumina焦磷酸测序法进行测序。使用BIPES、UCHIME、TSC和GAST分析样本中的物种信息。

结果

乳酸杆菌是健康女性中的优势菌(超过95%),主要包括惰性乳酸杆菌和卷曲乳酸杆菌,还有少量加德纳菌、颗粒链菌、链球菌、普雷沃菌、大肠杆菌等属。30例BV患者的α多样性显著增加(P<0.001),β多样性也明显改变,表现为乳酸杆菌减少(从45%降至1%,主要为惰性乳酸杆菌),甚至6例患者中乳酸杆菌缺失,加德纳菌、普雷沃菌、颗粒链菌、厌氧球菌、小单胞菌、哈氏消化链球菌、消化链球菌和戴阿李斯特菌的相对丰度增加。与之前的数据不同,7例BV病例显示稀有物种加氏乳酸杆菌和嗜酸乳酸杆菌占优势(75%至50%)。

结论

乳酸杆菌是健康女性阴道中的优势菌(主要是惰性乳酸杆菌和卷曲乳酸杆菌),与许多其他细菌和多种微生物共存。大多数BV患者中乳酸杆菌显著减少甚至缺失,部分病例显示稀有物种加氏乳酸杆菌和嗜酸乳酸杆菌占优势。

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