摘要
目的:利用DNA指纹图谱技术——变性梯度凝胶电泳(denaturing gradient gel electrophoresis,DGGE)分析我国育龄期及绝经期健康女性阴道菌群多样性,为了解各种下生殖道感染疾病状态下的微生态环境奠定基础,为研制适合恢复中国女性正常阴道微生态环境的微生态制剂提供依据。方法:选取2009年10月至2010年1月在北京大学第一医院妇产科门诊常规体检的健康女性(其中育龄期30例、绝经后30例)为研究对象。采集阴道分泌物,提取细菌总DNA,采用通用引物对细菌16S rDNA基因的V3区进行扩增,将PCR产物进行变性梯度凝胶电泳,通过对DGGE特征条带切胶回收并进行克隆测序,利用GenBank中的Blast与已知序列比对,鉴定细菌菌种,分析不同生理状态下阴道菌群的多样性。结果:(1)我国育龄期健康女性阴道常见菌种为:卷曲乳杆菌(Lactobacilluscrispatus)、惰性乳杆菌(Lactobacillus iners)和加氏乳杆菌(Lactobacillus gasseri),其中惰性乳杆菌是以往通过传统培养方法尚未认识到的阴道优势菌种。(2)绝经期健康女性阴道菌种与育龄期不同且复杂,常见菌种有:惰性乳杆菌、卷曲乳杆菌、大肠杆菌(Escherichia coli)、无乳链球菌(Streptococcus agalactiae)、解没食子酸链球菌(Streptococcusgallolyticus)、韦荣球菌属(Veillonella sp.)、中间链球菌(Streptococcus intermedius)、咽颊炎链球菌(Streptococcus angi-nosus)、普雷沃氏菌属(Prevotella sp.)、气球菌(Anaerococcus lactolyticus)和脆弱拟杆菌(Bacteroides fragilis)。结论:通过DGGE分析发现我国育龄期健康女性阴道常见优势菌种为:卷曲乳杆菌、惰性乳杆菌、加氏乳杆菌;绝经期健康女性阴道常见优势菌种为:惰性乳杆菌、卷曲乳杆菌、大肠杆菌、链球菌属、普雷沃菌属、脆弱拟杆菌以及产乳酸的韦荣球菌、气球菌属,并首次发现在不同生理状态下惰性乳杆菌均是阴道优势菌种之一。
Objective：To analyze the diversity of healthy women＇s vaginal dominant flora in different physiological states with PCR-denaturing gradient gel electrophoresis（PCR-DGGE）,to establish the basis for the future study of lower genital tract infections,and to provide a theoretical basis for the development of vaginal probiotics adapted to Chinese females.Methods： Women who underwent routine gynecologic examinations in Peking University First Hospital from October 2009 to January 2010 were enrolled,including 30 at reproductive age and 30 at post menopause age.Vaginal samples were collected and then total bacterial DNA was extracted.Universal bacterial primers were used to amplify the V3 region of 16S rDNA gene.PCR products were analyzed by denatured gradient gel electrophoresis（DGGE）.Featured bands on DGGE were recovered,cloned and sequenced.Alignment with known sequence was made by using Blast on GenBank to identify bacterial strains so as to analyze the diversity of healthy women＇s vaginal dominant flora in different physiological states.Results：（1） Vaginal flora in healthy Chinese women at their reproductive age were relatively simple,Lactobacillus were the dominant bacteria.Common bacteria included： Lactobacillus crispatus,Lactobacillus iners,and Lactobacillus gasseri.While L.iners were the dominant vaginal bacteria that could not be recognized by traditional method.（2） Vaginal bacteria in the women at post menopause age were more complex than in those at reproductive age.Common bacteria included： Lactobacillus iners,Lactobacillus crispatus,Escherichia coli,Streptococcus agalactiae,Streptococcus gallolyticus,Veillonella sp.,Streptococcus intermedius,Streptococcus anginosus,Prevotella sp.,Anaerococcus lactolyticus and Bacteroides fragilis.Conclusion： Based on the DGGE analysis,the most common vaginal bacteria in Chinese healthy women at reproductive age are Lactobacillus crispatus,Lactobacillus iners,and Lactobacillus gasseri.the most common vaginal bacteria in healthy post-menopausal
出处
《北京大学学报（医学版）》
CAS
CSCD
北大核心
2012年第2期281-287,共7页
Journal of Peking University:Health Sciences
基金
国家自然科学基金(81170535)资助~~
关键词
阴道
细菌
变性梯度凝胶电泳
女性
中国
Vagina
Bacteria
Denaturing gradient gel electrophoresis
Female
China
