通过特异PCR扩增和16S rDNA序列分析检测动弯杆菌

细菌性阴道病 (BacterialVaginosis,BV)是由于细菌过度生长所致阴道微生态非正常改变 ,从而导致的一类多微生物病 (PolymicrobialDiseases)。动弯杆菌 (Mobiluncussp .)与BV发生有密切关系 ,但该菌为厌氧菌 ,营养要求苛刻 ,很难进行纯培养 ,国内鲜有研究报道。本文先对BV动物模型恒河猴阴道分泌物进行厌氧菌混合培养 ,抽提混合物染色体DNA ,之后设计了一对动弯杆菌 16SrRNA基因的特异性引物 ,用PCR的方法扩增出了特异片段。通过对扩增产物进行测序分析 ,确定检测出的为动弯杆菌 ,并且与羞怯动弯杆菌极为相似     

细菌性阴道病的诊断和治疗

细菌性阴道病 (bacterial vaginosis,BV)是世界范围内常见的一种阴道疾病 ,该病与许多严重的妇产科并发症有直接联系 ,许多并发症可通过明确诊断并正确治疗 BV而得到了预防 [1 ,2 ]。本文就 BV诊断和治疗技术的研究进展作一综述。1　 BV的诊断BV是阴道微生态环境内菌群与分泌物生化性质变化引起的疾病 ,表现为正常阴道乳酸杆菌减少而代之以阴道加特纳菌、普雷沃菌、动弯杆菌、人型支原体等兼性厌氧、厌氧菌数量增加并伴随分泌物 p H值升高等重要生化成份变化的临床症候群。单一种微生物的定性培养结果并不能作为 BV诊断的准确依据。 1…     

动弯杆菌的研究进展

动弯杆菌是1984年命名的厌氧菌。该菌为条件致病菌,与细菌性阴道病密切相关。本文概述了动弯杆菌的主要生物学性状、致病性及实验室的检查方法。     

细菌性阴道病的诊断方法及评价

细菌性阴道病 ( Bacterial Vaginosis BV)是由于细菌过度生长所致阴道微生态的非正常改变 ,从而导致的一类多微生物病 ( Polymicrobial Diseases)。以前曾命名为非特异性阴道炎 ( nonspecific vaginitis) ,加特纳菌性阴道炎 ( Gardnerellavaginitis) ,厌氧菌性阴道炎 ( anaerobicvaaginosis) ,嗜血杆菌性阴道炎 ( Corynebecterium vaginitis) ,棒状杆菌阴道炎( H aemophilus vaginitis)等。直到 1984年 ,在瑞典国际会议上才正式命名为“细菌性阴道病”。 BV是最常见的妇产科疾病之一 ,通过对患有 BV的妇女的阴道分泌物进行培养后发现 …     

乳杆菌治疗细菌性阴道病的临床疗效观察

细菌性阴道病(BV)是妇科常见病,它是由于阴道内微生态平衡失调,伴有以下特征性症状:阴道内乳杆菌数量不足,伴需氧菌、厌氧菌和微需氧菌的混合性阴道菌丛以极大的数量过度生长,BV已被证实与盆腔炎和妊娠合并症相关,并有极高的复发率.该文以乳杆菌治疗298例细菌性阴道炎患者,观察评价其疗效.     

细菌性阴道的病因与治疗的研究进展

细菌性阴道病 ( Bacterial Vaginosis,BV)是妇科的常见病、多发病。它是由于阴道内微生态平衡失调 ,厌氧菌和阴道加德菌过盛生长 ,兼性厌氧性乳杆菌受抑制所引起的一种无阴道粘膜炎症性表现的综合征。患细菌性阴道病时 ,阴道内乳杆菌的数量比用药前低 1 0 0 0倍以上或完全消失。应用抗生素治疗 ,虽然可使 BV的症状得到暂时缓解 ,但也进一步地杀死本已少得可怜的乳杆菌 ,加重阴道微生态失调的程度 ,从而使 BV反复复发。由于 BV的主要病理生理改变是阴道菌群失调 ,因此恢复患者阴道微生态平衡 ,保持阴道的正常微生态环境的生态制剂 ,成为…     

阴道加德纳菌研究进展

细菌性阴道病(bacterial vaginosis,BV)主要由阴道加德纳氏菌(Gardnerella vaginalis,GV)与某些厌氧菌混合感染引起,伴有阴道分泌物性质改变的一组症候群,其病理特征无炎症病变和白细胞浸润。是广大妇女的常见病、多发病,占外阴阴道感染的30%~50%[1]。由于对BV病原菌认识的争议     

中国汉族育龄女性阴道内乳杆菌的鉴定与系统发育分析

目的比较基质辅助激光解吸电离飞行时间质谱(MALDITOF MS)与16S rRNA基因测序鉴定女性阴道乳杆菌方法,同时探究不同阴道微生态背景的汉族育龄女性阴道内乳杆菌的种类及其分布情况。方法对260例汉族育龄女性阴道拭子进行阴道微生态分析及乳杆菌的分离培养,根据阴道微生态鉴定结果分为正常组(173例)、中间型细菌性阴道病组(IBV,55例)、细菌性阴道病组(BV,9例)和外阴阴道炎假丝酵母菌病组(VVC,23例),分离乳杆菌并进行纯化培养,纯化的乳杆菌经MALDITOF MS和16S rRNA基因测序鉴定并进行聚类分析。结果2种方法鉴定的一致率为95.3%。260例标本中149例(57.3%)检出了乳杆菌,共发现17种乳杆菌,检出的乳杆菌分布于5个乳杆菌系统发育群,其中分离出较多的乳杆菌为卷曲乳杆菌(47例)、加氏乳杆菌(38例)、阴道乳杆菌(21例)。正常组116例(67.1%)标本分离出乳杆菌,IBV组19例(34.5%),BV组2例(22.2%),VVC组12例(52.2%)。正常育龄女性与患BV女性乳杆菌种类与数量差异有统计学意义(χ²=22.936 0,P<0.000 1),与患VVC女性差异无统计学意义(χ²=1.983 0,P=0.160 0)。与其他组相比,正常组育龄女性中分离出卷曲乳杆菌的比例显著高于BV组和VVC组(χ²=4.548 0,P=0.033 0;χ²=5.605 0,P=0.020 0),VVC组育龄女性分离出加氏乳杆菌的比例显著高于正常组(χ²=5.090 0,P=0.035 0)。结论对比16S rRNA基因测序与MALDITOF MS方法的乳杆菌鉴定结果,除了阴道乳杆菌和人阴道乳杆菌出现不一致的鉴定结果,其余15种乳杆菌鉴定结果一致性均为100%,但基于测序与基于质谱的方法产生的乳杆菌聚类分析结果不完全一致。卷曲乳杆菌可能是反映育龄女性正常阴道微生态的重要标志物,加氏乳杆菌可能与女性VVC的发生有关。     

乳杆菌活菌制剂对细菌性阴道病局部免疫的调节

目的检测正常人和细菌性阴道病(BV)患者治疗前后阴道局部细胞因子的变化,探讨乳杆菌活菌制剂对女性生殖道免疫的影响,为阴道微生态平衡与阴道黏膜免疫屏障的关系的研究提供一定的依据。方法用乳杆菌活菌制剂治疗BV,通过酶联免疫吸附试验法即ELISA法来检测BV患者治疗前后及正常健康妇女阴道局部细胞因子sIgA、IL-2、IL-13的水平。结果 sIgA、IL-13水平治疗前组较对照组明显升高(P<0.01),治疗后组较治疗前组水平下降(P<0.05),治疗后组较对照组水平升高(P>0.05);IL-2水平治疗前组较对照组明显降低,治疗后组较治疗前组明显升高(P<0.05),治疗后组较对照组下降(P<0.05)。结论 BV患者阴道局部免疫功能发生了改变;乳杆菌活菌制剂对BV患者阴道局部免疫具调节作用.     

细菌性阴道病肠道菌群与阴道菌群16S rDNA测序分析

目的通过对细菌性阴道病(BV)患者肠道菌群及阴道菌群16S rDNA扩增子测序,分析其结构、多样性、相关性以及BV对肠道菌群的影响,为今后治疗BV提供新的思路。方法选取符合纳入标准的BV患者11例(BV组),健康者9例(C组),留存阴道分泌物及新鲜粪便进行16S rDNA基因检测分析。结果C组阴道菌群以乳杆菌属为主,BV与加德纳菌属、普雷沃菌属、Sneathia、窄食单胞菌属(Stenotrophomonas)、阿托波菌属、Shuttleworthia、巨型球菌属密切相关。BV组肠道、阴道菌群丰富度均高于C组。Alpha多样性分析中C组和BV组肠道菌群、阴道菌群的Shannon指数组间比较,χ²值为29.137, P=0.000<0.05,两组阴道菌群Shannon指数组间比较差异具有统计学意义(P<0.05),BV组高于C组。物种多样性曲线反映本研究样本测序数据量的合理性,表明BV组的肠道菌群多样性、丰富度均高于C组,主坐标分析表明C组肠道与阴道的菌群结构差距较大,BV组肠道与阴道的菌群结构有相似之处,且两组肠道菌群结构接近。BV组阴道菌群中厚壁菌门丰度较C组低,放线菌门、拟杆菌门较C组高;BV组肠道菌群中拟杆菌门丰度较C组低;C组肠道中拟杆菌门明显高于阴道,厚壁菌门明显低于阴道; BV组阴道菌群中放线菌门丰度高于C组,差异均具有统计学意义(P<0.05)。结论BV阴道菌群与肠道菌群具有相关性,BV可能引起肠道菌群结构比例和多样性的改变。     

Bacterial communities of the coronal sulcus and distal urethra of adolescent males

Lactobacillus-dominated vaginal microbiotas are associated with reproductive health and STI resistance in women, whereas altered microbiotas are associated with bacterial vaginosis (BV), STI risk and poor reproductive outcomes. Putative vaginal taxa have been observed in male first-catch urine, urethral swab and coronal sulcus (CS) specimens but the significance of these observations is unclear. We used 16 S rRNA sequencing to characterize the microbiota of the CS and urine collected from 18 adolescent men over three consecutive months. CS microbiotas of most participants were more stable than their urine microbiotas and the composition of CS microbiotas were strongly influenced by circumcision. BV-associated taxa, including Atopobium, Megasphaera, Mobiluncus, Prevotella and Gemella, were detected in CS specimens from sexually experienced and inexperienced participants. In contrast, urine primarily contained taxa that were not abundant in CS specimens. Lactobacilllus and Streptococcus were major urine taxa but their abundance was inversely correlated. In contrast, Sneathia, Mycoplasma and Ureaplasma were only found in urine from sexually active participants. Thus, the CS and urine support stable and distinct bacterial communities. Finally, our results suggest that the penis and the urethra can be colonized by a variety of BV-associated taxa and that some of these colonizations result from partnered sexual activity.     

甲硝唑与乳杆菌活菌制剂对细菌性阴道病患者阴道微生物群影响的对比性研究

目的对300例细菌性阴道病（Bacterial Vaginosis,BV）患者随机使用甲硝唑（100例）、乳杆菌活菌制剂定君生（100例）及甲硝唑联合定君生（100例）,观察乳杆菌在阴道内定植情况及用药前后阴道内微生物多样性的变化,为临床治疗细菌性阴道病提供实验依据。方法分别提取300例BV患者用药前和用药后的阴道微生物总基因组DNA,以德氏乳杆菌16SrDNA特异性引物进行PCR,检测患者用药后德氏乳杆菌在阴道内的定植情况。以细菌16SrRNAV3区通用引物进行PCR,扩增产物进行变性梯度凝胶电泳（DGGE）,对所得指纹图谱进行分析,研究用药对微生态变化的影响状况。结果甲硝唑治疗组用药前后均没有德氏乳杆菌检出;定君生治疗组用药后德氏乳杆菌检出率为54％;甲硝唑联合定君生治疗组用药后德氏乳杆菌检出率为76％。甲硝唑治疗组用药后微生物多样性显著降低,微生物群结构失衡;定君生治疗组用药后微生物多样性有所降低,微生物群结构无显著差异;甲硝唑联合定君生治疗组用药后微生物多样性降低,但较前两组有所增加,微生物群结构差异无统计学意义。结论甲硝唑显著降低BV患者阴道微生物多样性,引起微生态失衡;定君生对BV患者阴道微生态的影响明显小于甲硝唑;定君生的使用有利于甲硝唑f预后阴道微生物多样性的恢复和微生态平衡的重建。     

Dynamics of vaginal bacterial communities in women developing bacterial vaginosis, candidiasis, or no infection, analyzed by PCR-denaturing gradient gel electrophoresis and real-time PCR

The microbial flora of the vagina plays a major role in preventing genital infections, including bacterial vaginosis (BV) and candidiasis (CA). An integrated approach based on PCR-denaturing gradient gel electrophoresis (PCR-DGGE) and real-time PCR was used to study the structure and dynamics of bacterial communities in vaginal fluids of healthy women and patients developing BV and CA. Universal eubacterial primers and Lactobacillus genus-specific primers, both targeted at 16S rRNA genes, were used in DGGE and real-time PCR analysis, respectively. The DGGE profiles revealed that the vaginal flora was dominated by Lactobacillus species under healthy conditions, whereas several potentially pathogenic bacteria were present in the flora of women with BV. Lactobacilli were the predominant bacterial population in the vagina for patients affected by CA, but changes in the composition of Lactobacillus species were observed. Real-time PCR analysis allowed the quantitative estimation of variations in lactobacilli associated with BV and CA diseases. A statistically significant decrease in the relative abundance of lactobacilli was found in vaginal fluids of patients with BV compared to the relative abundance of lactobacilli in the vaginal fluids of healthy women and patients with CA.     

健康妇女及3例细菌性阴道病患者阴道菌群的非培养分析

根据Amsel标准及Nugent标准,确诊筛选健康妇女及细菌性阴道病(bacterial vaginosis,BV)患者各3例.提取其阴道分泌物样本的总DNA,构建16S rRNA基因克隆文库,并对阳性克隆进行ARDRA和测序分析.结果表明,健康妇女样本的基因文库中,分别以卷曲乳酸杆菌(L.crispatus)和惰性乳酸杆菌(L. iners)的克隆子占较大比例,另外存在少量的阴道乳酸杆菌(L.vaginalis)或詹氏乳酸杆菌(L.jensenii)的克隆子.BV患者样本的基因文库中,克隆子所代表的菌种类型明显增多,但均以阴道加德纳氏菌(Gardnerella vaginalis)和阴道阿托波氏菌(Atopobium vaginae)的克隆子占较大比例,且无乳酸杆菌克隆子.说明健康妇女阴道菌群的种类单一,以乳酸杆菌占优势,L. iners为优势菌种之一;BV患者阴道菌群的种类复杂多样,但均以Gardnerella vaginalis及Atopobium vaginae共同占优势.     

健康妇女及3例细菌性阴道病患者阴道菌群的非培养分析

根据Amsel标准及Nugent标准, 确诊筛选健康妇女及细菌性阴道病(bacterial vaginosis, BV)患者各3例。提取其阴道分泌物样本的总DNA, 构建16S rRNA基因克隆文库, 并对阳性克隆进行ARDRA和测序分析。结果表明, 健康妇女样本的基因文库中, 分别以卷曲乳酸杆菌(L. crispatus)和惰性乳酸杆菌(L. iners)的克隆子占较大比例, 另外存在少量的阴道乳酸杆菌(L. vaginalis)或詹氏乳酸杆菌(L. jensenii)的克隆子。BV患者样本的基因文库中, 克隆子所代表的菌种类型明显增多, 但均以阴道加德纳氏菌(Gardnerella vaginalis)和阴道阿托波氏菌(Atopobium vaginae)的克隆子占较大比例, 且无乳酸杆菌克隆子。说明健康妇女阴道菌群的种类单一, 以乳酸杆菌占优势, L. iners为优势菌种之一; BV患者阴道菌群的种类复杂多样, 但均以Gardnerella vaginalis及Atopobium vaginae共同占优势。     

Dynamics of Vaginal Bacterial Communities in Women Developing Bacterial Vaginosis, Candidiasis, or No Infection, Analyzed by PCR-Denaturing Gradient Gel Electrophoresis and Real-Time PCR

The microbial flora of the vagina plays a major role in preventing genital infections, including bacterial vaginosis (BV) and candidiasis (CA). An integrated approach based on PCR-denaturing gradient gel electrophoresis (PCR-DGGE) and real-time PCR was used to study the structure and dynamics of bacterial communities in vaginal fluids of healthy women and patients developing BV and CA. Universal eubacterial primers and Lactobacillus genus-specific primers, both targeted at 16S rRNA genes, were used in DGGE and real-time PCR analysis, respectively. The DGGE profiles revealed that the vaginal flora was dominated by Lactobacillus species under healthy conditions, whereas several potentially pathogenic bacteria were present in the flora of women with BV. Lactobacilli were the predominant bacterial population in the vagina for patients affected by CA, but changes in the composition of Lactobacillus species were observed. Real-time PCR analysis allowed the quantitative estimation of variations in lactobacilli associated with BV and CA diseases. A statistically significant decrease in the relative abundance of lactobacilli was found in vaginal fluids of patients with BV compared to the relative abundance of lactobacilli in the vaginal fluids of healthy women and patients with CA.     

The evaluation of prevalence and the impact of pathological microflora of the lower genital tract among women at early pregnancy on the risk of preterm delivery

The main aim of this prospective study was to determine the prevalence and an association between pathological microflora of the lower genital tract diagnosed at early pregnancy and the risk of preterm delivery. The study group comprised 179 randomly selected pregnant women from Lodz region, between 8 and 16 week of pregnancy. For the qualitative and quantitative assessment of biocenosis of the lower genital tract vaginal and cervical swabs were collected from the pregnant women under study. The C. trachomatis antigen was detected by direct immunofluorescence assay. The vaginal swabs were tested for aerobic and anaerobic bacteria. Bacterial vaginosis was diagnosed by Gram stain according to Spiegel's criteria. To evaluate the risk factors odds ratios were calculated using EPI INFO software. 21 (11.7%) women delivered before 37th week of pregnancy. Bacterial vaginosis was diagnosed among 51 (28.5%) pregnant women while intermediate microflora was diagnosed by Gram stain in 62 (34.6%) women. The shortest mean gestational age at delivery was noted among women with BV. The rate of preterm delivery in BV group was 15.7% comparing to 9.1% among women with normal microflora. Among women with preterm delivery BV was diagnosed in 38.1% (OR = 1.86). Based on culture results only 84 (46.9%) women had normal microflora at early pregnancy. The pathological culture was associated with slightly increased preterm delivery rate (12.6%) as compare to 10.7% in control group. Positive culture for Bacteroides and Mobiluncus was connected with nonstatistical rise in the risk of preterm delivery. No association between C. trachomatis infection at early pregnancy and elevated risk of preterm delivery was found. Early pregnancy diagnosis of bacterial vaginosis and its treatment should lower the rate of prematurity in Poland.     

Transfer of members of the genus Falcivibrio to the genus Mobiluncus, and emended description of the genus Mobiluncus

It has long been thought that the genera Mobiluncus and Falcivibrio contain the same organisms. Using a polyphasic approach, it was found that Mobiluncus curtisii and Mobiluncus mulieris were the same as Falcivibrio vaginalis and Falcivibrio grandis, respectively. As the genus name Mobiluncus takes precedence, it is proposed that F. vaginalis and F. grandis be transferred to the genus Mobiluncus. In agreement with previous studies, results from phenotypic tests did not support the separation of M. curtisii strains into its two subspecies, M. curtisii subsp. curtisii and M. curtisii subsp. holmesii. Phenotypic complexity within M. curtisii dictates that the species should be treated as a complex until more in-depth analyses of the species have been performed. Phylogenetic analyses, based on 16S rRNA gene sequences, demonstrated that the genus Mobiluncus was associated with Varibaculum cambriense and the two subspecies of Actinomyces neuii, and that A. neuii is only distantly related to Actinomyces sensu stricto.