解脲支原体实验方法的探讨

本文选择了180例门诊泌尿生殖系感染患者,同时用PCR和培养法检测角脲支原体。其阳性率分别为31.1%,20.0%。培养法阳性率下降可能与临床用药有关,表明检测解脲支原体,PCR方法更占有优势,如为首诊病人,PCR结合培养更能提高解脲支原体感染的检测水平。     

男性生殖系感染者精液解脲支原体感染的检测研究

目的：探讨男性生殖系感染者精液解脲支原体（ＩＵＩ）感染的检测研究。方法：取精液标本检测，以源于解脲支原体尿素酶基因序列为引物，应用聚合酶链反应（ＰＣＲ）技术对解脲支原体ＤＮＡ进行扩增。结果：所试几种非解脲支原体培养物均阴性。解脲支原体阳性出现１条４１８ｂｐ的ＤＮＡ扩增带。３８２４例男性生殖道感染者，ＵＵ阳性检出率２９．１％。其中急性淋菌性生死道炎感染率为４８．１％，淋菌后生殖道炎感染率为３０．２％     

解脲支原体对胎儿输卵管粘膜上皮细胞的粘附作用

解脲支原体 (ＵｒｅａｐｌａｓｍａＵｒｅ ａｌｙｔｉｃｕｍ ,ＵＵ)是泌尿生殖道感染的重要致病因子 ,为了解ＵＵ是否对人输卵管粘膜上皮具有粘附作用 ,从而探讨ＵＵ对人输卵管粘膜的致病作用 ,采用人输卵管粘膜上皮体外粘附模型进行评价。选择因病引产的 6～ 7个月龄女性胎儿 ,而孕母无ＵＵ感染病史 ,孕期宫颈分泌物ＵＵ培养和ＵＵ ＤＮＡＰＣＲ检查均阴性 ,此次引产是因母体患有妊娠高血压综合征或心脏病等要求终止妊娠。在无菌的条件下取胎儿输卵管组织 (自宫角部切断 )共 8条 ,纵行剪开输卵管 ,暴露输卵管的粘膜面 ,并展平于木板上 ,用1…     

南充市门诊男性泌尿生殖道标本解脲支原体和沙眼衣原体的感染情况研究

目的分析南充市门诊就诊男性泌尿生殖道标本UU 和CT 感染情况以及年龄分布特点,加强本区域男性CT 和UU 筛查遥方法收集2014 年7 月~2017年7 月共657 例门诊男性尿道分泌物标本,采用荧光PCR 法进行UU尧CT测定,并将所有 男性受检者分为5 个年龄组,对所得检测结果进行统计学分析遥结果657 例受检者共有阳性267 例,总感染率40.64%,单一感 染率36.07%,单一CT 感染14.00%,UU感染22.07%,CT合并UU感染率4.57%遥不同年龄段中21~50 岁三个年龄组感染率最 高（分别44.28%尧41.18%尧43.90%）,21~30尧31~40尧41~50 岁三个年龄组分别与逸51 岁比较,差异有统计学意义（约0.05）遥此外, CT合并UU感染也主要分布在21~50 岁,所有年龄组UU感染阳性率高于CT 感染遥结论南充地区男性CT 和UU的感染 率均较高,以单一感染为主,尤其好发于21~50 岁遥加强CT 和UU 筛查,对性传播疾病防治尧泌尿生殖系感染的诊疗和优生优 育有重要意义遥     

光纤间接免疫荧光法检测肺炎支原体抗体

目的和方法:首次采用自行设计、研制的光纤荧光光度计对肺炎支原体抗体进行了检测,并将该法与临床上现行ELISA法及荧光显微镜法进行比较测定.结果:此法标准曲线的相关系数r=0.9930,测定的相对荧光强度(△F)大于2.0为阳性,小于2.0为阴性,且稳定性好.检测35人次临床血清标本,18例阳性标本中,本法有19例为阳性,与ELISA法及荧光显微镜法符合率(特异性)为94.4%;17例阴性标本中,本法有16例为阴性,符合率为94%结论:表明本光纤法具有特异性好、敏感性高,还具有可定量,操作简便快速,所需样本量微,所用仪器新颖、价廉、易普及等优点.     

应用PCR与培养法检测解脲支原体感染的研究

本文应用聚合酶链反应（ＰＣＲ）对１９８便病人进行检测，阳性率３８．９％，同时应用培养法进行对照检测，阳性率２１．７％。结果表明ＰＣＲ法检测 同于培养法检测阳性率，确定ＰＣＲ法是目前对支原体诊断的准确性、敏感性、特异上于最佳的检测方法。     

液体培养法检测育龄妇女生殖道感染标本中解脲支原体

目的了解育龄妇女生殖道解脲支原体感染情况。方法用液体培养法对2004年1月至2005年9月间拟诊为生殖道感染的年龄在23~40岁之间的育龄妇女宫颈分泌物标本检测解脲支原体。结果女性生殖道感染组解脲支原体阳性率为54.9%(282/514),高于对照组(22%);经卡方检验,二者之间有高度显著性差异(P<0.005)。结论解脲支原体与女性生殖道感染有关,育龄妇女生殖道解脲支原体感染率很高,并且逐年不断增高的趋势,所以育龄妇女应在孕前或产前检测解脲支原体,这对优生工作有重要意义。     

生殖支原体及解脲脲原体对妊娠的影响研究

目的：进一步探讨生殖支原体（Mg)及解脲脲原体（Uu)对妊娠的影响。方法：应用灵敏、特异的套式PCR（nPCR）技术对219例胎盘组织进行Mg、Uu检测,并对Mg、Uu nPCR阳性产物各1例进行了DNA序列测定以验证。结果：早产、胎膜早破者的Mg、Uu,胎儿窘迫者的Mg之阳性检出率均明显高于正常妊娠者（P均小于0．01）;另外,上胎流产、死胎者和本次妊娠3．00以上,呈强关联。结论：Mg与Uu一样,围产期感染可致不良妊娠结局。     

免疫组化法检测解脲脲原体的研究

目的　建立检测解脲脲原体 (UU)的实验方法。方法　应用亲和素 生物素 酶复合物技术 (ABC法 )观察UU感染者 ,并与培养法和PCR法相比较。结果　ABC法对 75例临床标本的检出率为 5 2 % ,与培养法和PCR法比较 ,在统计学上差异无显著性 (P >0 .0 5 )。结论　ABC法可作为诊断UU感染的快速检测方法 ,在临床应用中有一定实用价值。     

荧光定量PCR检测稽留流产患者沙眼衣原体和解脲支原体结果分析

目的探讨沙眼衣原体（CT）和解脲支原体（UU）感染与孕妇稽留流产的相关性,以及荧光定量PCR在稽留流产病因检测中的应用价值。方法采用荧光定量PCR检测稽留流产患者宫颈分泌物标本中的CT-DNA和UU-DNA的含量。结果 138例稽留流产患者中72例感染CT或UU,总检出率53.62%,其中CT阳性率21.74%,UU阳性率42.03%,混合感染10.14%。与正常人群对照差异均有统计学意义（P〈0.05）。结论泌尿生殖系统CT和UU的感染与稽留流产有密切相关性,荧光定量PCR对稽留流产患者CT和UU的检测简单、快速、准确。     

解脲脲原体套式（Nested）PCR检测研究

本文报告解脲脲原体（ＵＵ）的套式（ＮＥＳＴＥＤ）ＰＣＲ检测方法。经方法学考核表明，本法的特异性、灵敏度以及试剂的稳定性和对临检标本的顺应性均较好。９３份各种生殖道炎症患者之宫颈拭子标本套式ＰＣＲ检出２１份阳性（阳性率２２．６％），而市售ＰＣＲ试剂盒仅检出１份阳性（阳性率１．１％）。前者阳性检出率明显高于后者（Ｐ＜０．０１）。     

Ureaplasma urealyticum binds mannose-binding lectin

Mannose-binding C-type lectin (MBL) is an important component of innate immunity in mammals. Mannose-binding lectin (MBL), an acute phase protein, acts as an opsonin for phagocytosis and also activates the mannan-binding lectin complement pathway. It may play a particularly significant role during infancy before adequate specific protection can be provided by the adaptive immune system. Ureaplasma urealyticum has been linked to several diseases including pneumonia and chronic lung disease (CLD) in premature infants. We therefore investigated the ability of U. urealyticum to bind MBL. A guinea pig IgG anti-rabbit-MBL antiserum was produced. An immunoblot (dot-blot) assay done on nitrocellulose membrane determined that the anti-MBL antibody had specificity against both rabbit and human MBL. Pure cultures of U. urealyticum, serotype 3, were used to make slide preparations. The slides containing the organisms were then incubated with nonimmune rabbit serum containing MBL. Ureaplasma was shown to bind rabbit MBL with an immunocytochemical assay using the guinea pig IgG anti-rabbit MBL antiserum. Horseradish peroxidase (HRP)-labeled anti-guinea pig IgG was used to localize the reaction. The anti-MBL antiserum was also used in an immunocytochemical assay to localize U. urealyticum in histological sections of lungs from mice specifically infected with this organism. The same method also indicated binding of MBL by ureaplasma in human lung tissue obtained at autopsy from culture positive infants. Our results demonstrate that ureaplasma has the capacity to bind MBL. The absence of MBL may play a role in the predisposition of diseases related to this organism.     

聚合酶链反应检测丝虫病患者泌尿道沙眼衣原体及解脲支原体的研究

本文采用聚合酶链反应方法,检测了５０ 例丝虫病患者尿中沙眼衣原体（ＣＴ） 及解脲支原体（ＵＵ）ＤＮＡ,同时选择３０ 例正常人作为对照。结果显示,丝虫病患者尿中ＣＴＤＮＡ、ＵＵ ＤＮＡ 和两者混合感染检出率分别为３０ ％ 、２５ ％ 和２０ ％ ,高于正常对３６３％ （Ｐ＜０－０１）,ＣＴ及ＵＵ 感染与病人的病情、病程有一定的关联。提示乳糜尿的发生、发展与泌尿道ＣＴ 和ＵＵ感染有关。     

解脲脲原体生物膜胞外多糖的检测

目的 研究解脲脲原体(Ureaplasma urealyticum,Uu)4个型别标准株在体外形成的生物膜之胞外多糖的分布及结构成分.方法 将Uu标准株Parvo群中4、8血清型和T960群中3、14血清型进行体外生物膜培养后,扫描电镜下观察生物膜组成及结构,并在FITC-ConA/PI及ECA/PI双荧光染色后进行激光共聚焦显微镜观察及测定平均荧光强度.秩和检验及t检验分别比较两种荧光标记物、两生物群的总体平均荧光强度的差异.结果 4个型别Uu标准株均可在体外形成生物膜,生物膜结构主要呈网格状,胞外物质占大部分比例.在激光共聚焦显微镜下,Uu生物膜胞外多糖均可被FITC-ConA和ECA染色,FTTC-ConA呈网格状分布,ECA小片状聚集分布.FITC-ConA的总体平均荧光强度较ECA高,差异有统计学意义(P＜0.001).结论 Uu体外培养生物膜主要呈网格状结构,胞外多糖中含有葡萄糖、甘露糖、半乳糖、N-乙酰葡聚糖残基,并以葡萄糖、甘露糖残基为主.     

解脲脲原体生物被膜形成与耐药性的相关性研究

目的 研究解脲脲原体(Ureaplasma urealyticum,Uu)标准株及临床分离株体外形成生物被膜的能力及游离状态与形成生物被膜后药物敏感性的差异.方法 对Uu标准株3、8血清型(Uu3、Uu8)及从女性患者宫颈中分离鉴定的21株Uu临床株进行体外培养后,扫描电镜、激光共聚焦显微镜鉴定生物被膜形成,并在生物被膜形成前后进行约敏测定(四环素、红霉素、环丙沙星).配对秩和检验及x2检验分别比较Uu游离状态及形成生物被膜后最低抑菌浓度间及耐药率间的差异.结果 Uu3、Uu8及21株Uu临床株均具有体外形成生物被膜的能力.Uu形成牛物被膜后对四环素、红霉素及环丙沙星的最低抑菌浓度较游离状态明显增高(P<0.001).Uu形成生物被膜后对红霉素及环丙沙星的耐药率增高具有统计学意义(P值分别为<0.001及0.035),但对四环素的耐药率增高无统计学意义(P=0.293).结论 Uu标准株及临床株均具有体外形成生物被膜的能力,Uu形成生物被膜后对抗菌素的抵抗力增加,出现了多重耐药现象.

Abstract：

Objective To study the ability of standard strain and clinical isolates of Ureaplasma spp. to form biofilms in vitro and to compare the antibiotic susceptibility of sessile cells and their planktonic counterparts. Methods A total of 21 Ureaplasma wealyticum(Uu) isolates recovered from female patients diagnosed with cervicitis and Uu serovar 3 and Uu serovar 8( Uu3, Uu8) were included. Scanning electron microscope and confocal scanning laser microscopy were used to identify biofilm formation. Conventional antibiotic susceptibility tests and biofilm susceptibility assays for tetracycline, erythromycin and ciprofloxacin were carried out. The paired rank sum test and was applied to analyze the statistical differences between the MIC and the minimal biofilm inhibitory concentration. The x2 test was applied to analyze the statistical differences of global resistance percentages between planktonic cells and sessile cells. Results Uu3, Uu8 and 21 Uu isolates all can form biofilms in vitro. Minimal inhibitory concentration of sessile cells compared with planktonic cells were obviously higher for tetracycline, erythromycin and ciprofloxacin (P <0.001). Global resistance percentages between planktonic cells and sessile cells were different for erythromycin (9.52% vs 61.90% , P < 0. 001), ciprofloxacin ( 80. 95% vs 100% , P = 0. 035 ) and tetracycline (4. 76% vs 14.29% , P =0.293). Conclusion Uu isolates and Uu1, Uu8 all can form biofilms in vitro, and biofilm formation can strengthen resistance of Uu to antibiotics, even multidrug resistance was observed.     

Analysis of the reactivity of indirect immunofluorescence in patients with pemphigus foliaceus and pemphigus vulgaris using rat bladder epithelium as a substrate

OBJECTIVES:

To evaluate the reactivity of indirect immunofluorescence using rat bladder epithelium as a substrate in patients with pemphigus foliaceus and pemphigus vulgaris from the Department of Dermatology, University of São Paulo Medical School, Brazil.

METHODS:

Thirty-two patients (8 male and 24 female) from the Department of Dermatology, University of São Paulo Medical School, were selected. Three had mucosal pemphigus vulgaris, 20 had mucocutaneous pemphigus vulgaris, and 9 had pemphigus foliaceus. Patients'' sera were tested by indirect immunofluorescence performed on human foreskin and rat bladder epithelium and by ELISA assays utilizing baculovirus-expressed recombinant desmoglein 3 and desmoglein 1.

RESULTS:

No patients with mucosal pemphigus vulgaris, 5 of 20 patients with mucocutaneous pemphigus vulgaris (25%) and 4 of 9 patients with pemphigus foliaceus (44%) had positive indirect immunofluorescence using rat bladder epithelium as a substrate.

CONCLUSION:

Indirect immunofluorescence using rat bladder epithelium as a substrate is recommended whenever a diagnosis of paraneoplastic pemphigus is considered. The identification of a subset of pemphigus foliaceus and pemphigus vulgaris patients that recognizes desmoplakins by this laboratory tool is critical to avoid the misdiagnosis of paraneoplastic pemphigus.     

解脲脲原体感染的检测与体外耐药性分析

目的探讨本地区解脲脲原体(Uu)感染及耐药情况,指导临床合理用药。方法对临床送检的531份标本进行Uu培养、计数、鉴定和药敏试验;对775份标本进行荧光定量PCR(FQ-PCR)法和培养法检测Uu。结果FQ-PCR法检出Uu阳性339例,阳性率43.74%。培养法检出Uu206例,阳性率38.79%。Uu对9种抗菌药物的耐药率最高为环丙沙星(CIP)80.10%,最低为交沙霉素(JOS)0.00%,敏感率最高为原始霉素(PRI)98.54%。结论Uu用两种方法检测均可有效检出。培养法能提供药敏结果,更有利于临床治疗。药敏结果提示本地区Uu感染经验用药可选择交沙霉素和强力霉素。