Evaluation of patients with dry eye disease for conjunctival Chlamydia trachomatis and Ureaplasma urealyticum

AIM：To determine the possibility of the development of dry eye disease（DED） as a result of persistent infection with Chlamydia trachomatis and Ureaplasma urealyticum in the conjunctiva of patients.METHODS： This study was conducted on 58 patients of age range 20-50 y,diagnosed with DED confirmed by Schirmer I test and tear breakup time.The non-dry eye control group included 27 subjects of the same age.Ocular specimens were collected as conjunctival scrapings and swabs divided into three groups： the first used for bacterial culture,the second and third taken to detect Chlamydia trachomatis and Ureaplasma urealyticum by direct fluorescent antibody（DFA） assay and polymerase chain reaction（PCR） method. RESULTS： Chlamydia trachomatis was detected in 65.5% and 76% of DED patients by DFA and PCR methods respectively.Ureaplasma urealyticum was found in 44.8% of DED infected patients using the PCR method.Both organisms were identified in only 37.9% of DED patients found to be infected.Control subjects had a 22%detection rate of Chlamydia trachomatis by DFA assay versus a 7% detection rate by PCR; while Ureaplasma urealyticum was detected in 3.7% of the controls by PCR method.The conjunctival culture revealed that gram positive microorganisms represented 75% of isolates with coagulase negative Staphylococci the most common（50%） followed by Staphylococcus aureus（20%）,whereas gram negative microorganisms occurred in 25% of cases,isolating Moraxella spp.as the most frequent organism. CONCLUSION： Our results tend to point out that Chlamydia trachomatis and Ureaplasma urealyticum were detected in a moderate percentage of patients with DED,and could be a fair possibility for its development.PCR is more reliable in detecting Chlamydia trachomatis than DFA technique.The presence of isolated conjunctival bacterial microflora can be of some potential value.

Study of the prevalence and association of ocular chlamydial conjunctivitis in women with genital infection by Chlamydia trachomatis, Mycoplasma genitalium and Candida albicans attending outpatient clinic

AIMTo determine the association between chlamydial conjunctivitis and genital infection by Chlamydia trachomatis, Mycoplasma genitalium and Candida albicans, in addition to the possible relationship between cultured bacterial pathogens and oculogenital chlamydial infection.METHODSThis study was performed on 100 (50 symptomatic and 50 asymptomatic) women attending the Gynecological and Obstetric outpatient clinic of Alzahra hospital, Alazhar University. Simultaneously a conjunctival swab was taken from these patients. Polymerase chain reaction (PCR) was done on DNA extracted from both vaginal and conjunctival swab samples. Culture for both vaginal and conjunctival swabs was also done.RESULTSCandida albicans was the predominant organism isolated by culture in 20% and 40% of conjunctival and vaginal swabs respectively. By the PCR method, ocular Chlamydia trachomatis was present in 60% of symptomatic women, while genital Chlamydia trachomatis infection was present in 30% of symptomatic women. The results of this method also indicated that 25/50 (50%) vaginal swabs were positive with PCR for Candida albicans versus 15/50 (30%) were PCR positive in conjunctival swab. Mycoplasma genitalium was present in only 10% of vaginal swabs. Concomitant oculogenital PCR positive results for Chlamydia trachomatis and Candida albicans were 30% and 28% respectively.CONCLUSIONOcular Chlamydia trachomatis was associated with genital Chlamydia trachomatis in a high percentage of women followed by Candida albicans. Cultured bacterial organisms do not play a role in enhancement of Chlamydia trachomatis infection.

Detection of <i>Chlamydia trachomatis</i>and <i>Neisseria gonorrhoeae</i>in Egyptian Women Suffering from Infertility

Chlamydial and gonococcal infections are recognized as two of the major causes of sexually transmissible human bacterial infection which may lead to infertility. In this cross sectional study, we aimed to determine the prevalence of Neisseria gonorrhoeae, Chlamydia trachomatis among Egyptian women using different microbiological methods. One hundred and fifty cervical swabs were collected, of which 100 were from infertile women. Culture and ELISA technique were used for screening of Neisseria gonorrhoeae and Chlamydia trachomatis individually. In addition, PCR was used for all examined samples. For C. trachomatis, 3 cases were positive for antigen detection by ELISA. Moreover, in obtained results of PCR, DNA was detected in 4 samples, and three of them from infertile group. So based on PCR results, the sensitivity and specificity of ELISA were 75% and 100% respectively. Furthermore, 3 samples were positive for gonococcal infections by PCR, and two of them were taken from infertile women. Positive results of two samples were verified by culture. The estimated sensitivity and specificity of culture method were 66.7% and 100% respectively. Results of this study indicate that PCR is a valuable method for detection of gonococcal and chlamydial infection and it is suitable for the confirmation of ELISA results for C. trachomatis diagnosis. Culture method is less sensitive than PCR for detection of N. gonorrhoeae. The prevalence of such infections is higher among infertile women.

Detection of Genitourinary Tract Chlamydia trachomatis Infection in Urine specimens by PCR Assay

Objective: To compare the sensitivity and specificity of thecervical/urethral swabs with voided urine specimens for thedetection of genitourinary tract infection with Chlamydia tra-chomatis and determine whether urine specimens can replacethe cervical/urethral swabs in detection of C. thachomatis. Methods: The matched cervical/urethral swabs and voidedurine specimens were collected from 569 patients of STD clinics.Polymerase chain reaction (PCR) assay specific for C. tra-chomatis plasmid DNA and rapid antigen testing (Clear viewassay) was used to detect C trachomatis. Standard criteria thatdefined 'true' positive included: 1) positive PCR results both incervical/urethral swab and voided urine specimen or 2) positivevoided urine results both by PCR assay and clear view test or 3)positive results in both PCR assay of cervical/urethral swab andclear view test of voided urine. For statistical analysis, thechi-square test was used. Results: The prevalence of C trachomatis in patients withsymptoms was 12.1% (28/231) in women and 10.4%(10/96) inmen, with no significant difference between them (x^2=0.21,P>0.05). The prevalence of C trachomatis in patients with nosymptoms was 11.0% (11/100) in women and 15.5% (22/142) inmen, with a significant difference existing between them. (x^2=4.0, P<0.05). No significant difference (P>0.05) existed betweenPCR testing of swabs (sensitivity 87.3%; specificity 99.2%) andPCR testing of urine (sensitivity 88.7%: specificity 98.8%). Asfor clear view assay, sensitivity was 60.6% and specificity was100%. Conclusions: PCR assay is superior to clear view in detectingC trachomatis. Although both PCR testing of swabs and PCRtesting of urine specimens both have high sensitivity and speci-ficity, urine specimen testing is more cost-effective, practical andnoninvasive. Thus urine specimens can take the place of theswabs in PCR testing for chlamydia.

Study on vertical transmision of Chlamydia trachomatis using PCR and DNA sequencing

Objective To investigate the vertical transmission rate of Chlamy dia trachomatis (CT) in Chongqing, China. Methods Specimens taken from 278 women and from their 79 infants were e xamined b y cell culture, polymerase chain reaction (PCR) and DNA sequence analysis. Chlam ydia trachomatis was isolated in McCoy cell culture. CT DNA was extracted with a modified NaI method. After cloning, recombinant plasmids were used for sequence analysis with the dideoxy chain termination method.Results 10.8% (30/278) of the cervical cultures of pregnant women were positive for Chlamydia trachomatis, while the positive rate tested by PCR was 14.0% (39/2 78). The vertical transmission rate of Chlamydia trachomatis was 55. 0% (11/20). The incidences of conjunctivitis and pneumonia in infants with Chlamydia t rachom atis positive mothers were 27.3% and 18.2%, respectively. DNA sequence s of Chlam ydia trachomatis isolated from the cervix of a mother and the nasopharynx of her baby were identical.Conclusion Chlamydia trachomatis infection is quite common in Cho ngqing , China. Our report is the first report of CT vertical transmission proved by DN A sequence analysis.

Detection of Chlamydia pneumonia DNA in nasopharyngolaryngeal swab samples from patients with rhinitis and pharyngolaryngitis with polymerase chain reaction

Objective To assess the prevalence of Chlamydia pneumomia DNA in patients with otolaryngic disease Methods PCR assay was used to detect Chlamydia pneumonia specific Pst Ⅰ 474 fragment DNA in swabs from patients with acute or subacute pharyngolaryngitis or rhinitis and sinusitis C pneumonia specific antibodies in sera were also assayed with microimmuno fluoresence (MIF) Results About 28% (49/175) of the patients were PCR positive and 25 7%(45/175) were MIF antibodies positive The accordance rate of the two methods was 91 8% Conclusion It is suggested that the C pneumonia infection was common in this group of patients and the C pneumonia Pst Ⅰ474 specific PCR was sensitive and specific for detecting C pneumonia in pharyngolaryngitis or rhinitis and sinusitis

Relationship between Chlamydia pneumoniae infection and occurrence of bronchial asthma

Objective: To study the relationship between Chlamydia pneumoniae (C. pneumoniae) infection and asthma exacerbation. Methods: A prospective study of C. pneumoniae infection was conducted in 75 patients with asthma and 63 patients with respiratory tract infection, and 100 blood donors served as controls. The presence of infection was convinced by the polymerase chain reaction and direct immunofluorescence assay for C. pneumoniae DNA from throat swab specimens and micro-immunofluorescence testing for C. pneu-moniae-specific IgG, IgM and IgA antibodies. Results: Prevalence of specific IgG in asthma patients (81. 3%) was higher than that of the blood donors (68. 0%, P<0. 05) and was not significantly different from respiratory tract infection patients (68. 0%, P>0. 05). The acute C. pneumoniae infection rate of symptomatic asthma patients (59. 4%) was markedly higher than that of respiratory tract infection patients (34. 9% , P<0. 05). The average titer of C. pneumoniae IgG instead of IgA in asthma patients (48. 38±6. 94) was significantly higher than respiratory tract infection patients (24. 70±8. 77, P<0. 05). Other pathogens were identified in 12 of 21 (57. 1%) asthma patients with C. pneumoniae. The symptoms of 7 asthma patients with C. pneumoniae infection were improved through antibiotic treatment. Conclusion: The findings suggest a possible role of C. pneumoniae infection in asthma.

不孕妇女宫颈沙眼衣原体感染的临床观察

目的 :探讨宫颈沙眼衣原体 (CT)感染对生育的影响。方法 :应用聚合酶链反应 (PCR)技术 ,对 2 16对不孕夫妇宫颈分泌物、尿道分泌物进行 CT- DNA检测 ,并观察宫颈 CT感染对围排卵期宫颈粘液葡萄糖含量及性交后试验的影响。结果 :2 16对不孕夫妇衣原体感染的阳性率分别为 :女性 32 .4 % ,男性 30 .1%。宫颈 CT感染妇女围排卵期性交后试验与未感染妇女比较 ,每高倍视野的活精数明显减少 (P<0 .0 5 ) ,围排卵期宫颈粘液葡萄糖含量明显减少 (P<0 .0 1)。经治疗后宫颈 CT转阴的妇女 ,围排卵期性交后试验每高倍视野的活精数明显高于治疗前 (P<0 .0 5 ) ,宫颈粘液的葡萄糖含量亦明显高于治疗前 (P<0 .0 1)。治疗后宫颈 CT持续阳性妇女 ,围排卵期性交后试验与治疗前比较 ,每高倍视野的活精数无明显变化 (P>0 .0 5 ) ,宫颈粘液的葡萄糖含量亦无明显变化 (P>0 .0 5 )。结论 :宫颈 CT- DNA感染是不孕的一个重要因素 ,导致不孕的原因 ,可能与患者宫颈粘液葡萄糖含量降低 。

195例妇女生殖道沙眼衣原体检出率分组比较研究

作者建立了以聚合酶链反应检测沙眼衣原体的方法，并以此技术对４组妇女生殖道沙眼衣原体感染进行了检测。结果表明卖淫妇女、ＳＴＤ门诊患者、妇科门诊患者及产科门诊孕妇生殖道沙眼衣原体的检出率分别为２４．４％（１１／４５）、１０．３％（５／４９）、６．０％（３／５０）及４．０％（２／５１）。前两组性病高危妇女沙眼衣原体的检出率（１７．０％）明显高于后两组妇女的检出率（５．０％）（ｐ＜０．０５），说明生殖道沙眼衣原体感染与性乱呈正相关。

PCR检验技术在性病临床标本实验诊断中应用价值的研究

目的 评价聚合酶链反应 (PCR)检测技术在性病临床诊断中的应用价值。方法 比较PCR与传统检测手段对淋病和非淋菌性尿道炎病原体的检测结果。淋球菌以加多粘菌素的GC血液琼脂分离 ;衣原体用McCoy细胞培养 ;支原体用高效支原体培养基培养。PCR试验用基因释放剂作前处理 ,以 1.5 %琼脂糖凝胶电泳观察结果。结果 用PCR法检查淋病、衣原体和支原体感染的临床标本其敏感性和特异性均比传统的培养法高 ,作PCR试验的标本取材要求不如作培养那样严格 ,但作PCR应使用正规的试剂盒、规范的操作和严格的防污染措施 ,否则极易引起交叉污染。结论 PCR检验技术在性病诊断中具有巨大的应用潜力 ,对一些培养困难的病原菌 ,可在规范操作和标准化试剂盒的基础上逐步加以使用。PCR检测不宜用作判愈。

新型MGB探针在沙眼衣原体实时PCR检测中的应用

为建立基于TaqMan MGB探针的沙眼衣原体DNA荧光定量PCR检测方法 ,探讨其临床应用价值 ,用PCR法扩增沙眼衣原体隐蔽质粒pLVG44 0 2 464～ 2 980nt段 ,并克隆入 pMD18 T载体用作参比模板 ,设计一对引物和一个TaqMan MGB探针 ,优化反应条件 ,建立沙眼衣原体DNA荧光定量PCR检测系统 ,并运用该系统同时应用连接酶链式反应 (LCR)法对临床标本进行检测 .结果显示所建立的沙眼衣原体DNA荧光定量PCR检测系统 ,最低检测限度为 1DNA拷贝每反应 ;在 10 0 ～ 10 9DNA拷贝每反应范围内 ,Ct 值 (每个反应管内的荧光信号达到设定的域值时所经历的循环数 )和DNA拷贝数呈线性关系 (r >0 990 ) ;对临床标本检测结果同LCR分析结果吻合率为 10 0 % .以上结果表明 ,所建立的基于TaqMan MGB探针的沙眼衣原体DNA荧光定量PCR检测系统具有敏感性高、特异性强和线性检测范围广等特点 。

泌尿生殖系感染患者尿液中沙眼衣原体、淋球菌、解脲脲原体感染分析

目的检测泌尿生殖系统感染患者尿液中沙眼衣原体（CT）、淋球菌（NG）、解脲脲原体（UU）感染状况,并进一步了解CT、NG、UU病原体在深圳市的感染情况,为临床诊治提供实验室依据。方法采集疑似泌尿系统感染的609例患者（其中男305例,女304例）尿液,并选取其中108例患者同时采集尿液、分泌物,采用实时荧光聚合酶链反应（PCR）检测CT、NG、UU。结果 CT、NG、UU在分泌物及尿液中的检测结果比较差异无统计学意义（P〉0.05）。单纯感染UU阳性率最高,为42.04%（256/609）;CT＋NG＋UU感染患者感染率为0.33%（2/609）;CT＋UU感染是双重感染中最高,为2.63%（16/609）。男性患者CT、NG、UU感染率分别为：29.18%（89/305）、14.10%（43/305）、52.45%（160/305）;女性患者CT、NG、UU感染率分别是：3.29%（10/304）、2.63%（8/304）、31.58%（96/304）。〈20岁者CT、NG、UU阳性检出率为64.00%,20~29岁者为63.16%,〉29~39岁者为63.34%,〉39岁者为75.00%。结论采用尿液和分泌物检测CT、NG、UU的检出率基本一致;CT、NG、UU的感染率后者明显高于前二者,混合感染以CT＋UU感染率最高;男性患者CT、NG、UU感染率均高于女性。

连接酶链反应检测宫颈沙眼衣原体感染

为评价连接酶链反应(LCR)诊断宫颈沙眼衣原体(CT)感染的意义，用质粒LCR和聚合酶链反应(PCR)检测170例STD门诊女性就诊者宫颈拭子标本中的CT，对两项检测结果不一致的标本进行校正，对PCR阳性而LCR阴性者，将LCR标本稀释10倍重复LCR或用PCR反应的DNA模板作LCR检测。对PCR阴性而LCR阳性者，用另一对针对沙眼衣原体主要外膜蛋白(MOMP)基因的引物作PCR测试。将以上两项检测结果阳性的标本判断为真阳性，确定LCR和PCR的敏感性和特异性。发现170例患者中24例LCR检测阳性(14.2％)，26例PCR阳性(15.3％)，对8例两项结果不一致的标本作了校正。LCR检测的敏感性和特异性分别为92％，99.3％；而PCR分别为92％，97.9％。结果提示LCR诊断女性宫颈CT感染具有较高的敏感性和特异性。

四川东北地区女性生殖道感染沙眼衣原体、淋球菌和解脲脲原体的结果分析

目的分析川东北地区2005~2012年女性生殖道沙眼衣原体(CT)、淋球菌(NG)和解脲脲原体(UU)感染状况及变化趋势,为该类患者的诊断和治疗提供实验室依据。方法应用实时荧光定量PCR(RT-PCR)技术对1 386例女性生殖道标本进行CT、NG、UU病原体DNA检测,并对结果进行统计分析。结果 3种病原体的总阳性率为62.8%(871/1 386)。单纯感染中以UU阳性率最高,为48.0%(665/1 386);CT、NG阳性率为仅为2.2%。混合感染以CT+UU阳性率最高,为6.5%(90/1 386);UU+NG阳性率为2.5%(35/1 386);CT+NG阳性率为0.4%(5/1 386);CT+UU+NG阳性率为1.1%(15/1 386)。不同年龄段中,小于20岁组阳性率为49.3%,而大于20岁组中均超过60%。2005~2012年间,女性CT、NG、UU病原体阳性率持续高水平且总体呈上升趋势。结论 CT和UU是该地区女性生殖道感染的主要病原体,并以20岁以上生殖道感染者阳性率均较高,3种病原体感染率呈逐年上升趋势,应该重视该人群CT和UU的检测以指导临床医师对患者的诊断和治疗。

盆腔粘连与沙眼衣原体和解脲支原体感染的关系

目的 探讨沙眼衣原体 (chlamydiatrachomatis,CT)和解脲支原体 (ureaplasmaurealyticum ,UU)感染与盆腔粘连的关系及其发病机制 ,并试图找到预防和降低盆腔粘连发生的方法和途径。方法 分别采用聚合酶链反应 (PCR)技术、金标免疫斑点法 ,将 5 0例妇科既往无盆腔手术史 ,行腹腔镜和开腹手术中有盆腔粘连患者作为盆腔粘连组 (观察组 ) ,4 0例同期妇科腹腔镜和开腹手术中无盆腔粘连患者为对照组。测定宫颈分泌物和手术标本中的CTDNA、UUDNA ,以及两组患者血清中抗CT和UU抗体。结果 观察组宫颈分泌物CT和UUDNA阳性率分别为 32 %和 4 0 % ;血清CT和UU抗体阳性率分别为 34%和 4 8% ;手术标本CT和UUDNA阳性率分别为 2 6 %和 30 %。对照组宫颈分泌物CT和UUDNA阳性率分别为 7 5 %和 1% ;血清CT和UU抗体阳性率分别为 5 %和 10 2 5 % ;手术标本CT和UUDNA阳性率分别为 2 5 %和 5 %。两组比较差异有显著性 (P <0 0 1)。结论 沙眼衣原体、解脲支原体感染与盆腔粘连有着密切关系。

沙眼衣原体垂直传播基因水平的快速检测

应用细胞培养法对86名孕妇及其新生儿进行沙眼衣原体(CT)的分离,然后采用聚合酶链反应(PCR)结合限制性片段长度多态性(RFLP)分析法,对CT阳性的母婴标本进行基因型鉴定。结果显示,86例孕妇CT阳性20例,阳性孕妇的新生儿有8例感染,垂直传播率40.0％。8对母婴相对标本鉴定后证实每对母婴感染的CT基因型一致。提示CT有很高的垂直传播率,应早期进行孕期筛查,降低新生儿CT感染率。

6932例患者泌尿生殖道沙眼衣原体与淋病奈瑟菌感染调查分析

目的了解泉州地区人群泌尿生殖道沙眼衣原体(CT)和淋病奈瑟菌(NG)临床感染现状,为临床诊断治疗提供参考。方法选取2019年9月至2020年3月在该院泌尿科、妇产科、皮肤性病科就诊的患者6932例,其中男性患者1320例,女性患者5612例。采用荧光定量聚合酶链反应检测患者泌尿生殖道的CT和NG。结果6932例患者CT和NG阳性率分别为8.98%和6.85%,二者同时检出患者达1.60%。男性患者CT和NG阳性率分别为3.48%和4.95%,二者同时检出患者占1.11%;女性患者CT和NG阳性率分别为5.50%和1.90%,二者同时检出患者占0.49%。男性NG感染率(4.95%)显著高于女性(1.90%),差异有统计学意义(P<0.05);而男性CT感染率(3.48%)低于女性(5.50%),差异有统计学意义(P<0.05)。从年龄分布来看,以20~30岁年龄段2种病原体感染率最高。结论泉州地区泌尿生殖道CT、NG感染率不尽相同,该地区男性患者NG感染率高于女性,而女性患者CT感染率高于男性,CT、NG同时感染在男性患者中较为常见,20~30岁年龄段感染率最高,应该加强监测。

肺炎衣原体感染与小儿哮喘发作

为了解肺炎衣原体 (Cpn)感染与小儿哮喘之间的关系 ,并探讨哮喘患儿Cpn感染的诊断及治疗 ,采用微量免疫荧光(MIF)试验及巢式聚合酶链式反应 (nPCR)2种方法对45例哮喘发作期患儿及20例正常对照组进行Cpn检测。结果 :①哮喘组14例nPCR检测阳性 ,对照组1例阳性 ,两者相比差异有显著性 (P<0.05) ,表明Cpn感染与小儿哮喘发作密切相关。②14例nPCR检测阳性患儿在随后的发作中5例 (35.71 %)仍为阳性 ,表明哮喘患儿中慢性或重复Cpn感染较常见。③就诊或住院后5例Cpn感染哮喘患儿接受了大环内酯类抗生素治疗 ,其中3例病情缓解 ,表明对Cpn感染哮喘患儿进行抗Cpn治疗是必要的。④经全自动荧光测序 ,随机抽取的2例阳性标本和Cpn标准株 (CWL_29)的nPCR产物DNA序列完全一致。

糖原测定法诊断女性生殖道沙眼衣原体感染

用糖原测定法诊断生殖道沙眼衣原体(Ct)感染。用蒽酮法检测女性生殖道上皮细胞内糖原的含量,并与诊断沙眼衣原体感染的聚合酶链反应(PCR)法作比较。320例宫颈标本的检测结果与 PCR 比较,糖原测定法诊断 Ct感染的敏感性为84.8%,特异性为96.5%.糖原测定法是一种简便、快速的诊断 Ct 感染的方法,适宜基层医疗单位采用。

335例儿童肺炎衣原体急性下呼吸道感染分析

目的 了解肺炎衣原体在急性下呼吸道感染儿童中的感染状况。方法 采用套式聚合酶链反应技术 ,对335例急性下呼吸道感染儿童的咽拭子标本进行了肺炎衣原体检测。结果 在 335例急性下呼吸道感染儿童中 ,共检测出肺炎衣原体阳性标本 2 5例 ,阳性率为 7.4 6 %。