阴道加德纳菌生物型与细菌性阴道病的关系

对１２０例细菌性阴道病患者和９０例正常对照组进行阴道加德纳菌的分离鉴定,同时采用Ｐｉｏｔ分型法对分离出的９３株Ｇｖ进行生物分型,结果显示ＢＶ患者Ｇｖ的检出率（５９．２％）明显高于正常对照组（２４．４％）,Ｐ〈０．０１。９３株Ｇｖ中,８个生物型全检测到,其中１型２３．７％,２型１８．３％,４型３２．３％,６型１０．８％,３型和５型均为５．４％,７型和８型为２．２％。ＢＶ患者主要为１、２、４型,显著高于正常对照组（８７．３％和３１．８％,Ｐ〈０．０１）。     

阴道液中唾液酸酶活性、阴道加德纳菌及其IgA水平的相关性研究

目的探讨阴道液中唾液酸酶活性、阴道加德纳菌及抗阴道加德纳菌的溶血素(anti-Gvh)IgA水平在细菌性阴道病(BV)的相关性。方法对15例健康人和60例临床诊断为BV的患者,取阴道分泌物分别进行唾液酸酶活性和anti-Gvh IgA水平测定及阴道加德纳菌(Gv)分离培养。唾液酸酶活性利用从底物-5溴-4氯-3吲哚基-α-D-N乙酰基神经氨酸(X-Neu5Ac)转化而得到的甲氧基苯酚的纳摩尔数来表示;anti-Gvh IgA通过ELISA法测定。结果BV组Gv活菌数(6.96 log CFU/g)显著高于健康对照组(2.58 log CFU/g)(P<0.01)。BV组anti-GvhIgA(238.0±220.6)显著高于健康对照组(175.0±40.16)(P<0.01)。BV组Gv分离率(86.7%)显著高于健康对照组(33.3%)(P<0.01)。45例临床诊断为BV的患者中,其中26例Gvh IgA(+),19例Gvh IgA(-);Gvh IgA(-)组阴道液中唾液酸酶活性(5.00±1.29)显著高于Gvh IgA(+)组(1.58±1.22)(P<0.01),2组的Gv的分离率和活菌数差异却没有显著性(P>0.05)。结论BV患者阴道内高活性的唾液酸酶与低水平anti-Gvh IgA和黏膜IgA破坏程度具有关联性。     

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     

利用X-Neu5Ac为底物测定唾液酸酶活性在诊断细菌性阴道病的价值

目的利用5溴-4氯-3吲哚乙酰基神经氨酸盐（X-Neu5Ac）为底物测定阴道唾液酸酶活性诊断细菌性阴道病（bacterial vaginosis,BV）的价值.方法健康妇女30例,临床Amsel法诊断为BV的患者45例,共计75例进行了阴道分泌物分析和检测,并与唾液酸酶活性法诊断作了对比研究.取阴道分泌物作为标本分别进行唾液酸酶活性和阴道菌群定量分析,检测细菌包括乳酸杆菌、类杆菌、肠杆菌、葡萄球菌、肠球菌和阴道加德纳菌.唾液酸酶活性测定利用的底物为X-Neu5Ac,特异活性用其产物 ——甲氧基苯酚的纳摩尔数来表示.结果阴道液唾液酸酶活性测定诊断细菌性阴道病的敏感性、特异性、阳性预期值和阴性预期值分别为88.9%、90%、93%和84.3%.唾液酸酶法在检测细菌性阴道病上和传统的Amsel法比较,差异无显著性（P＞0.05）.唾液酸酶阳性组Gv活菌数（6.96 log CFU/g）明显高于唾液酸酶阴性组（2.05 log CFU/g）（P＜0.01）.唾液酸酶阳性组产H2O2阴道乳杆菌（LB＋）活菌数（4.26 Log CFU/g）明显低于唾液酸酶阴性组（8.66 Log CFU/g）（P＜0.01）.唾液酸酶阳性组与唾液酸酶阴性组两组的阴道液中需氧菌活菌数差异无显著性（P＞0.05）,主要包括金黄色葡萄球菌、肠球菌和肠杆菌.结论利用X-Neu5Ac作为唾液酸酶的底物测定唾液酸酶活性的方法是诊断细菌性阴道病的有效检测方法.阴道内唾液酸酶活性增强,厌氧菌数量增加,LB＋数量减少,提示BV发生恶化.     

492例细菌性阴道病患者加德纳菌检测与药敏分析

目的了解当地女性细菌性阴道病(BV)患者中阴道加德纳菌(GV)的感染情况以及GV对常用抗菌药物的敏感性,为BV的诊断和治疗提供依据。方法对收集的492例BV患者和151例正常健康妇女的阴道分泌物进行GV的分离培养与鉴定,并对分离菌进行常用抗菌药物敏感性试验。结果 492例BV患者GV阳性率为27.8%,健康妇女GV阳性率为7.9%,两者比较差异有统计学意义(P<0.05)。分离的GV对万古霉素和克林霉素均较敏感,对头孢噻肟、四环素、左氧氟沙星、氨苄青霉素、红霉素、甲硝唑的敏感性依次降低。结论 GV为当地BV患者的重要病原菌之一,且对万古霉素和克林霉素均较敏感,对甲硝唑的敏感性最低。     

中药安菌凝胶对狐狸阴道加德纳菌病的实验研究

通过对狐狸加德纳菌病（ＧＶＦ）阴道菌群分析、阴道光镜和电镜下组织学结构观察认定,ＧＶＦ与人类细菌性道病（ＢＶ）同属菌群失调所致,并以此为动物模型,采用黄芪、苦参、蛇床子、甘草四味中药以新型高分子材料卡波姆（Ｃａｒｂｏｍｅｒ）为赋形剂制成凝胶,对其进行体外抑菌实验和不同组方的疗效比较,结果显示：Ｂ组方中药制剂（安菌凝胶）治疗后的疗效最好,关在细胞水平上中药作为微生态调节剂的机制进行了初步探讨。     

治疗前后加德纳菌载量 对细菌性阴道病患者复发风险的预测价值

目的评价治疗前后加德纳菌载量预测细菌性阴道病(bacterial vaginosis,BV)患者复发风险的临床价值。方法将2017年1月至2018年1月于我院就诊的312例BV患者纳入研究,根据治疗后BV是否复发分为复发组(114例)及未复发组(198例),分别比较两组患者治疗前后阴道分泌物的pH值、Nugent评分、乳杆菌数量和加德纳菌载量,并应用受试者工作曲线(ROC)对上述指标预测BV复发的诊断价值及诊断效能进行评价。结果治疗前及治疗后复发组患者的pH值(t=2.187、3.822,均P0.05)、Nugent评分(t=1.976、5.611,均P0.05)、加德纳菌载量(t=5.089、16.204,均P0.05)均明显高于未复发组,乳杆菌数量(t=8.245、8.899;均P0.05)明显低于未复发组;多因素Logistic回归分析显示治疗后阴道分泌物Nugent评分及加德纳菌载量均是BV复发的独立危险因素,而乳杆菌数量为保护因素;对上述指标预测BV复发的相关性进行分析,ROC曲线显示,加德纳菌载量预测BV复发的AUC为0.892,明显高于pH值、Nugent评分、乳杆菌数量(AUC=0.580、0.648、0.772),诊断最佳截点为1.761×10~5 copy/mL,此时的敏感性和特异性分别为93.5%和75.4%。结论 BV患者治疗后的加德纳菌载量在预测其复发方面具有较高的敏感性及特异性,可用于预测BV复发。     

新的人兽共患传染病——狐狸阴道加德纳氏菌病的研究 Ⅳ.Dot-ELISA检测狐狸阴道加德纳氏菌病血清抗体的研究

应用建立的Dot-ELISA方法检测了192份狐狸阴道加德纳氏菌人工感染狐、疫苗免疫狐和菌检阳性狐血清,结果187/192呈阳性反应,检出率为97.3%. 阻断和交叉试验证实,本方法具有特异性.敏感度为平板凝集试验(PAT)的28.1倍,为微量凝集试验(MAT)的5.5倍.重复性良好,与PAT、MAT的阳性符合率为100%.     

新的人兽共患传染病——狐狸阴道加德纳氏菌病的研究 Ⅳ.Dot-ELISA检测狐狸阴道加德纳氏菌病血清抗体的研究

应用建立的Dot-ELISA方法检测了192份狐狸阴道加德纳氏菌人工感染狐、疫苗免疫狐和菌检阳性狐血清,结果187/192呈阳性反应,检出率为97.3%. 阻断和交叉试验证实,本方法具有特异性.敏感度为平板凝集试验(PAT)的28.1倍,为微量凝集试验(MAT)的5.5倍.重复性良好,与PAT、MAT的阳性符合率为100%.     

一种新的人兽共患传染病——狐狸阴道加德纳氏菌病的研究：VI.阴道加德纳氏茵病灭活疫苗的研制

选用免疫原性优良的我国狐狸阴道加德纳氏菌主要流行型血清Ｉ型ＧＶＦ４４号菌株,先后试验研制了氢氧化铝胶、蜂胶和油佐剂灭活苗。试验结果表明,铝胶苗的安全性优,免疫剂量为４０亿菌／１ｍｌ,３倍免疫剂量的铝胶灭活苗接种狐狸后无任何不良反应,免疫后２１ｄ经１００个ＩＤ５０强毒的攻击,对Ｉ型的保护率达９２％,对Ⅱ、Ⅲ型的保护率达８０％以上。免疫持续期为６个月,４￣１０℃条件下可保存１０个月。通过田间试验应用证     

Loop‐mediated isothermal amplification for the rapid detection of Gardnerella vaginalis

The aim of this study was to develop a method for the rapid detection of Gardnerella vaginalis, which is proposed to play a key role in the pathogenesis of bacterial vaginosis. Specific loop‐mediated isothermal amplification (LAMP) primers were designed and used to detect target DNA within 45 min under isothermal conditions. Comparative screening indicated that the LAMP assay is superior to PCR in terms of rapidity, and is equivalent in sensitivity and specificity. This LAMP assay can be used for rapid screening and detection of G. vaginalis in vaginal samples; the limit of detection is 10 fg DNA.

     

Physical characterization of the pore forming cytolysine from Gardnerella vaginalis

Abstract The cytolytic toxin (CTox) produced by Gardnerella vaginalis is able to form voltage-dependent cationic channels when incorporated in lipid membranes (Moran et al, 1991) FEBS Lett. 283, 317–320). Osmotic protection experiments show that toxin incorporated in human erythrocytes forms pores between 18 Å and 28 Å in diameter. A hypothesis of pore formation as a primary event to produce cytolysis is proposed. The CTox activity increases when cells are depolarized by increasing the extracellular K⁺ concentration, probably reflecting the voltage dependent character of CTox formed channels. The cytolytic effect of the toxin was prevented by low temperatures and was a function of the extracellular Ca²⁺ concentration, suggesting a Ca²⁺ influx as part of the lytic mechanism. Binding of CTox to erythrocytes was dependent on external Ca²⁺ and was less temperature-dependent. Dose-response analysis suggests cooperativity of the toxin for the lytic activity, although no direct evidence of oligomerization has been found.     

Lactobacillus plantarum Lp62 exerts probiotic effects against Gardnerella vaginalis ATCC 49154 in bacterial vaginosis

The severe side-effects elicited by conventional antibiotic therapy and the recurrence of Bacterial vaginosis-associated bacteria and bacterial resistance have led to the development of novel alternative therapies, among which genital probiotics are widely used. In this study, we aimed to evaluate the antimicrobial activities of Lactobacillus plantarum Lp62 and its supernatant against Gardnerella vaginalis, using both in vitro and in vivo approaches. In vitro assays were used to evaluate the viability of the strain and the antimicrobial activities of the supernatant in different pH ranges. An in vivo assay was performed on female BALB/c mice, wherein the animals were divided into eight groups: four control groups and four treated groups (for curative and preventive therapies). After infecting and treating the mice, the animals were killed to quantify the bacterial load using qPCR, evaluate leucocyte cellular response, determine vaginal cytokine levels and perform cytokine tissue gene expression. Our analyses revealed significant activity of the strain and its supernatant against G. vaginalis. Preliminary in vitro tests showed that the strain grew with equal efficiency in different pH ranges. Meanwhile, the presence of halo and inhibition of pathogen growth established the significant activity of the supernatant against G. vaginalis. We observed that both micro-organisms are resident bacteria of mouse microbiota and that the lactobacilli population growth was affected by G. vaginalis and vice versa. We also observed that the treated groups, with their low bacterial load, absence of leucocyte recruitment, reduced cytokine levels in the vaginal lavage and normalized cytokine gene expression, successfully controlled the infection.     

Lactobacillus strains isolated from the vaginal microbiota of healthy women inhibit Prevotella bivia and Gardnerella vaginalis in coculture and cell culture

The purpose of this study was to investigate how human vaginal isolates of Lactobacillus acidophilus, Lactobacillus jensenii, Lactobacillus gasseri and Lactobacillus crispatus inhibit the vaginosis-associated pathogens Gardnerella vaginalis and Prevotella bivia. Results show that all the strains in coculture condition reduced the viability of G. vaginalis and P. bivia, but with differing degrees of efficacy. The treatment of G. vaginalis- and P. bivia-infected cultured human cervix epithelial HeLa cells with L. gasseri strain KS120.1 culture or cell-free culture supernatant (CFCS) results in the killing of the pathogens that are adhering to the cells. The mechanism of the killing activity is not attributable to low pH and the presence of lactic acid alone, but rather to the presence of hydrogen peroxide and proteolytic enzyme-resistant compound(s) present in the CFCSs. In addition, coculture of G. vaginalis or P. bivia with L. gasseri KS120.1 culture or KS120.1 bacteria results in inhibition of the adhesion of the pathogens onto HeLa cells.     

细菌性阴道病联合测定技术在BV诊断中的应用

目的 探讨生化法在细菌性阴道病(BV)诊断中的应用价值.方法 分别使用镜检法和生化法对1466例就诊者的阴道分泌物进行检测.结果 在1 466例受检者中,使用生化法检出BV患者1 297例,使用镜检法检出BV患者1 301例.经统计学分析,二者检出BV的阳性率差异无统计学意义(x2=0.46,P＞0.05),且二者的符合率为97.14％ (1424/1466).结论 生化法检测准确快速,适宜在基层医院推广使用.     

2930例应用两种实验方法诊断细菌性阴道炎的比较

目的探讨细菌性阴道炎（BV）实验方法在临床的应用。方法对2009年3月至2009年8月在我院门诊就诊的患者2930例,随机分为2组,采用生理盐水直接涂片镜检法与细菌性阴道病联合测定试剂盒检测法进行比较。结果直接涂片镜检法阳性率为22.03%;应用细菌性阴道病联合测定试剂盒检测法,阳性率为45.81%,其中单患BV组占22.20%,合并白细胞酯酶2个加号占7.70%,合并白细胞酯酶3个加号占16.08%。结论细菌性阴道炎合并感染时,运用细菌性阴道病联合测定试剂盒检测法进行阴道分泌物检查,检出率明显高于涂片镜检法。