【作者】 朱艳宾；

【导师】 薛凤霞；

【作者基本信息】 天津医科大学 ， 妇产科学， 2013， 博士

【摘要】 近年研究发现,引起阴道细菌感染的疾病有两类,一类由厌氧菌和兼性厌氧菌引起,临床上称为细菌性阴道病(BV),另一类由需氧菌引起,临床上称需氧菌性阴道炎(AV)。关于BV的研究很多,而关于AV的研究国内报道较少,妊娠期AV以及对妊娠结局的影响国内尚未见报道,国外报道不多。B族链球菌(GBS)是AV常见致病菌,妊娠期GBS感染可导致严重的不良妊娠结局,但母婴GBS的血清型及基因型配对研究国内报道较少。因此,我们选取在我院产科门诊建册常规产检、诊断为需氧菌性阴道炎(AV)的孕妇150例为研究对象,分析阴道菌群分布情况及其与妊娠结局相关性,并对其母婴配对GBS菌株血清型及基因型进行了相关性分析。第一部分妊娠期需氧菌性阴道炎对妊娠结局的影响目的探讨AV孕妇妊娠不同时期阴道菌群分布情况及其与妊娠结局相关性。方法选取2010年7月～2012年7月在深圳市南山人民医院产科门诊建册常规产检、诊断为AV的孕妇150例为研究对象,分为三组,其中早孕组50例(孕周<13周),中孕组50例(13周≤孕周<28周),晚孕组50例(孕周≥28周),并选取同期在我院产科门诊行健康检查的正常孕妇100例为对照组。取阴道分泌物,部分镜检,部分送微生物培养,AV阳性者根据患者意愿给予治疗。分析AV孕妇阴道菌群分布,并随访阴道分泌物、妊娠结局包括分娩孕周、分娩方式、胎膜破裂、绒毛膜羊膜炎、胎儿窘迫、产褥感染、产后出血、新生儿评分、新生儿体重及新生儿感染情况。结果1.AV孕妇以阴道分泌物性状改变就诊者占31.3%(47/150),分泌物一般为黄色或黄绿色、稀薄、脓性。阴道菌群主要以GBS、大肠埃希氏菌、金黄色葡萄球菌及粪肠球菌为主,其中GBS检出率为28.6%(43/150),早孕组32%,中孕组28%,晚孕组26%。2.150例AV孕妇胎膜早破(26%)、绒毛膜羊膜炎(10.7%)、产后出血(9.3%)的发生率均明显高于对照组的发生率(分别为13.0%、2.0%、3.0%)(P<0.05)；各个孕期AV孕妇胎膜早破、绒毛膜羊膜炎、产后出血发生率差异无统计学意义(P>0.05)。3.AV孕妇所分娩的新生儿中,新生儿感染率为11.3%,高于对照组的2%(P<0.05)；而新生儿体重、Apgar评分明显低于对照组(P<0.05)；共发生新生儿肺炎4例,其中AV孕妇分娩新生儿3例,均为GBS感染。4.应用克林霉素磷酸酯阴道泡腾片治疗后,治疗组胎膜早破(20.6%)、绒毛膜羊膜炎(4.9%)、产后出血(5.9%)的发生率明显低于未治疗组孕妇的发生率(分别为37.5%、22.9%、14.6%)(P<0.05)。而分娩方式、胎儿窘迫、早产、产褥感染发生率两组比较,差异均无统计学意义(P>0.05)。5.治疗组新生儿感染率为8.9%,明显低于未治疗组的16.7%(P<0.05)；而新生儿体重、Apgar评分明显高于未治疗组(P<0.05)。结论AV感染孕妇主要症状为阴道分泌物呈黄色、黄绿色、稀薄、脓性,感染菌群主要以GBS、大肠埃希氏菌、金黄色葡萄球菌及粪肠球菌为主,AV感染可导致胎膜早破、绒毛膜羊膜炎、产后出血、新生儿感染、窒息、低出生体重等不良妊娠结局,经治疗可改善妊娠结局。第二部分母婴感染GBS血清型及基因型分析目的探讨母婴配对GBS菌株血清型和DNA指纹图谱是否一致。方法选取第一部分AV中为GBS感染者及其新生儿口咽部分泌物培养G1BS阳性者为研究对象,随机选取GBS菌株72株利用免疫双扩散法进行血清学分型,并将母婴菌株配对,研究其血清型是否一致,应用RAPD技术对18株不同血清型的菌株进行DNA指纹图谱研究,比较G1BS菌株血清型和基因型是否一致。结果1.对43例GBS感染者进行药敏试验,GBS对青霉素G、氨苄青霉素、头孢唑啉、头孢替安及克林霉素敏感,敏感率均为97.7%,红霉素和头孢哌酮的耐药率较高,红霉素耐药率高达23.3%。2.对72株GBS菌株进行了血清型分析,共分离出9个血清型,包括Type Ⅰ。、Ⅰb、IⅡ、Ⅲ、Ⅳ、Ⅴ、Ⅵ、Ⅶ、Ⅷ,其中Ⅱ型占23.6%,Ⅲ型占25%,是主要血清型,另外发现3株GBS为不可分型。3.利用免疫双扩散法将12例新生儿GBS供试抗原与其母亲GBS抗血清进行配对检测,12例均产生沉淀线,相应的沉淀线完全融合,形成平滑的曲线。4.应用RAPD技术对18株GBS基因型进行研究,所用7条随机引物中有4条呈现良好的多态性和稳定性,可产生差异显著的指纹图谱。18株GBS间的遗传距离为0.082～0.531,平均距离为0.35,其中第4株和第13株同为血清型TypeⅢ,但遗传距离最大,为0.358；第1株和第7株为不同血清型,而遗传距离最小,为0.011。不同血清型的GBS菌株具有相似的指纹图,而同一血清型的菌株指纹图存在差异。结论1.我们共分离出9个血清型,其中Ⅱ型和Ⅲ是主要血清型,另外发现3株GBS为不可分型,孕妇及其产婴儿GBS菌株具较高的相似性。2.不同血清型的GBS菌株具有相似的指纹图,同一血清型的菌株指纹图存在差异。更多还原

【Abstract】 Aerobic vaginitis is common vaginitis of women.Aerobic vaginitis in pregnant women can seriously affect the quality of perinatal care, cause many adverse effects on pregnancy. Studies have found that group B streptococcus is common vaginitis pathogen of aerobic vaginitis, but aerobic bacteria during pregnancy on pregnancy outcome and gene effects and maternal and child group B Streptococcus serotype type is not reported in China. Therefore, we collected150cases of different maternal during the2010-2012in our hospital of vaginal secretions, analysis of flora distribution and pregnancy outcome and correlation, and the mother-baby pairs and gene of GBS strain serotype type.Part I The relationship between aerobic vaginitis in pregnant women and pregnant outcomeObjective To investigate the vaginal flora distribution and its correlation with pregnancy outcomes of aerobic vaginitis women of pregnancy in different periods.Methods We choiced150cases of aerobic vaginitis in pregnant women from July2010to July2012in our hospital routine prenatal obstetric outpatient service.50cases of first trimester group (gestational age<13weeks),50cases of second trimester group (13weeks<gestational age<28weeks),50cases of third trimester group (gestational age≥28weeks). And we also choiced100cases of normal pregnant women during the same period in the hospital routine prenatal obstetric outpatient service as control group. Vaginal Secretion Smearing and Culturing. Then we followed-up pregnant outcome including gestational weeks, the Mode of delivery, rupture of membranes, chorioamnionitis, puerperal infection, fetal distress, postpartum hemorrhage, neonatal weight, Apgar score and infection in neonates.Results1. The mainly bacteria in aerobic bacterium vaginitis infection of pregnant women were group B streptococcus, e. coli bacteria, staphylococcus ball bacteria and enterococcus. There was no statistically significant difference between each pregnancy group(P>0.05). 2.There was statistically significant difference between aerobic bacterium vaginitis women and normal pregnant women in incidence of premature rupture of membrances(P<0.05);There was no statistically significant difference among different aerobic bacterium vaginitis pregnant women in the incidence of premature rupture of membranes(P>0.05). There was statistically significant difference between aerobic bacterium vaginitis women and normal pregnant women in incidence of chorioamnionitis (P<0.05); There was no statistically significant difference among different aerobic bacterium vaginitis pregnant women in incidence of chorioamnionitis (P>0.05). There was statistically significant difference between aerobic bacterium vaginitis women and normal pregnant women in the incidence of postpartum hemorrhage(P<0.05); There was no statistically significant difference among different aerobic bacterium vaginitis pregnant women in incidence of postpartum hemorrhage (P>0.05).3. There was statistically significant difference between aerobic bacterium vaginitis women and normal pregnant women in the incidence of and infection in neonates(P<0.05);. neonatal weight, Apgar score of newborn of aerobic bacterium vaginitis women were lower than normal pregnant women(P<0.05).4. After treatment,There was statistically significant difference between the treatment group and non-treatment group in incidence of premature rupture of membranes(P<0.05).In pregnant women, The bacterial culture of aerobe incidence of chorioamnionitis of102cases of treatment group was4.9%, lower than incidence of chorioamnionitis of non-treatment pregnant women and the difference was statistically significant (P<0.05). The non-treatment pregnant women had significantly higher incidence of postpartum hemorrhage than treatment women(P<0.05). while the incidence of fetal distress and preterm birth and puerperal infection in the two groups, there were no statistically significant difference (P>0.05).5. There was statistically significant difference between untreated pregnant women and treatment group in the incidence of infection in neonates(P<0.05); neonatal weight, Apgar score of newborn of untreated pregnant women were lower than treatment group (P<0.05). Conclusion1. The mainly bacteria in aerobic bacterium vaginitis infection of pregnant women were group B streptococcus, e. coli bacteria, staphylococcus ball bacteria and enterococcus. There was no statistically significant difference between each pregnancy group.2. Aerobic bacterium vaginitis can cause many adverse effects such as:premature rupture of fetal membrane,chorioamnionitis,postpartum hemorrhage. After treatment, Pregnant outcome woulde be improved.Part Ⅱ:Analysis of serotype and Genotype of Group B streptococcus in pregnant women and their newbornsObjective To investigate mother son paired GBS serotype strains, and DNA fingerprint are consistent, in order to be early prevention, early treatment, improve the outcome.Methods We selected of vaginal secretion culture for group B streptococcus of pregnant women and their newborns oropharyngeal secretions training GBS as the research object, in selecting strains, used immune double diffusion method and compare serum type of credits and mother son paired GBS serotype strains. Through RAPD, We studied GBS genotype.Results1. We used immune double diffusion method to analyze serotype GBS bacteria of72strains.9serotype were isolated, including Ⅱ and Ⅲ main serotype, another three GBS inseparable type strains.2.12cases of neonatal GBS antigen and its matching mother GBS antiserum precipitation line corresponding to precipitate line total integration formed a smooth curve. It showed that they had the same both GBS main antigen components, a quite close relationship, namely the strains with high similarity.3. Through RAPD, We studied of18kinds of GBS genotype.7random primers can be used in four of the article presents good polymorphism and stability, and can produce significant difference of the fingerprint. To analysis the genetic distance between the different strains.18strains of GBS genetic distance were frome0.082to0.531, the average distance of0.35,18strains of group B streptococcus4strains and13strains of genetic distance, the biggest0.358.1and7of the smallest genetic distance was0.011. GBS of different serotype strains had similar fingerprint, the same serotype strains had different fingerprint.Conclusion1. A total of nine serotype was isolated, of which Ⅱ and Ⅲ were main serum types. We also found that three strains of GBS were inseparable type. GBS production strains between pregnant women and their babies were high similarity.2. GBS of the same serotype strains had different fingerprint,different serotype strains had similar fingerprint.更多还原