生殖支原体和解脲脲原体与非淋菌性尿道炎相关性的研究

目的:研究生殖支原体(Mg)及解脲脲原体(Uu)与男性急性非淋菌性尿道炎(NGU)的相关性。方法:采用病例对照研究,收集性病门诊NGU病人、性病门诊有非婚性行为但无尿道炎的就诊者及健康体检者三组人群的尿道拭子,用AMPLICORCTNGPCRTest进行沙眼衣原体(Ct)的检测,巢式PCR(nPCR)和培养法进行Uu、人型支原体(Mh)的检测,nPCR及扩增产物DNA测序进行Mg的检测。结果:三组人群中Mg的检出情况分别为:25例(25%)、6例(6.4%)和0(P<0.0001),其中6例NGU病人为Ct与Mg混合感染。在NGU和门诊对照组中,Mg阳性者罹患急性NGU的危险性是Mg阴性者的3.917倍,(95%CI:1.618～9.126);在Ct阴性的NGU(NCNGU)中,Mg感染使罹患急性NGU的危险性增加7.2倍(95%CI:2.9～18.0)。nPCR检出三组人群中Uu的携带者分别为:40例(40%)、42例(44.7%)和46例(46.9%)。结论:Mg感染与男性急性NGU呈强相关,Uu与NGU无相关性。     

不同人群宫颈分泌物中沙眼衣原体感染状况分析

目的探讨不同人群宫颈分泌物中沙眼衣原体检测的意义。方法采用免疫层析法对来自妇科门诊、孕前检查门诊和性病门诊的6320例女性进行宫颈分泌物中沙眼衣原体检测。结果在妇科门诊、孕前检查门诊和性病门诊受检人群中沙眼衣原体阳性率分别为3.78%(153/4048),4.03%(87/2160),56.68%(59/112),性病门诊受检者的衣原体阳性率明显高于其他两组人群。结论衣原体检测在妇科门诊、孕前检查门诊和性病门诊中具有十分重要的价值,对提高妇女生殖健康水平、促进优生优育和防止性传播疾病蔓延起到积极的作用。     

男性无症状性病高危人群尿沉渣检测沙眼衣原体

目的　研究能否用尿沉渣代替尿道拭子检测沙眼衣原体 (Ct)。方法　分别用Wellcozymechlamydia法和PCR法对 35 2例男性无症状的性病高危人群首段尿沉渣进行Ct检测 ,两法经尿沉渣、尿道拭子配对研究。结果　以尿道拭子Ct培养为金标准 ,两种方法的敏感性和特异性分别为 5 0 98% (P <0 0 5 )和 10 0 % (P >0 0 5 ) ,98 0 4% (P >0 0 5 )和 96 6 8% (P >0 0 5 )。结论　对Ct的检测 ,用Wellcozymechlamydia法时 ,不宜用尿沉渣代替尿道拭子作标本 ;而用PCR法时 ,可用尿沉渣代替尿道拭子作标本。     

应用PCR试验的DNA提纯试剂盒来处理Gen-Probe沙眼衣原体PACE2试验的标本

以核酸扩增试验为基础的确证试验可以增加沙眼衣原体筛查试验的敏感性和特异性。Gen-Probe杂交试验是最常用的筛查试验之一。然而,迄今尚无将PCR确证试验与Gen-Probe杂交试验联用的报道。作者以Roche Amplicor PCR沙眼衣原体试验用于对Gen-Probe PACE 2     

聚合酶链反应检测男性非淋菌性尿道炎沙眼衣原体和解脲支原体

应用聚合酶链反应(PCR)检测了139例男性非淋菌性尿道炎(NGU)患者泌尿道标本中的沙眼衣原体(CT)和解脲支原体(UU)的DNA,结果:沙眼衣原体的阳性率为53.96％(75例),解脲支原体的阳性率为19.43％(27例),其中仅1例患者二种病原体同时阳性。结果表明沙眼衣原体和解脲支原体是男性非淋菌性尿道炎的主要致病菌;对检测阴性的患者,有必要做进一步的检测,以利于临床有效治疗。     

临床检测沙眼衣原体方法的实用性探讨

沙眼衣原体(CT)是非淋菌性尿道炎(NGU)、宫颈炎的病原体之一,约占NGU病原体的30%～50%.1实验室确诊依据为检测沙眼衣原体抗原.目前其检测方法较多,但灵敏度和特异性相差较大.我们对95例有典型症状和病史的NGU患者用胶体金法和多聚酶链式反应(PCR)两种方法同时检测沙眼衣原体,比较两种方法的实用性.结果报道如下.     

沙眼衣原体检测室间质量评价标本的保存时间探讨

拭子标本具有易保存及运送携带方便等优点,因此,被广泛应用于质控标本（品）的制作.我们亦采用拭子作为开展沙眼衣原体（Ct）检测室间质量评价（室间质评）标本的制作.为保证质控标本的质量,本实验室对2006年10月制备的Ct质控标本进行了为期1年的检测观察,以探讨标本可保存的最佳时间.本文将英国Unipath公司生产的Ct快速免疫层析试剂与美国Roche公司生产的Amplicor 衣原体PCR试剂用于质控标本的检测,现将结果报道如下.     

沙眼衣原体培养、LCR与荧光PCR在检测泌尿生殖道沙眼衣原体的意义

为了比较细胞培养、连接酶链反应（LCR）和荧光PCR技术在检测性传播疾病门诊患者标本沙眼衣原体的意义。分别在国内5家临床医院性传播疾病门诊收集到673份尿道／宫颈拭子标本，进行沙眼衣原体培养和荧光PCR检测，检测结果不相符合的标本采用LCR复检，比较分析PCR与培养、LCR以及综合结果的相符性。结果合格病例616例，培养和荧光PCDR的检测阳性率分别为6．33％和27．1％，与培养相比，荧光PCR法的敏感性为100％。LCR复核标本200份，与之相比，荧光PCR的敏感性为98．6％，特异性89．5％，YI指数为0．881。综合分析证明荧光PCR检测沙眼衣原体的敏感性为98．78％，特异性为98．67％，YI指数0．9745。结果临床表现发现，男性患者出现尿道炎症体征意义大于症状，而在女性患者症状或体征均不特异。结果表明国产荧光PCR技术检测尿道／宫颈拭子沙眼衣原体具有较高的敏感性与特异性，可以用于临床试验， 控与监督是本方法得以正确应用的关键。     

应用PCR联合检测500例泌尿生殖道感染三种性病病原体

淋球菌(NG)、沙眼衣原体(Ct)和解脲支原体(Uu)混合感染越来越引起人们重视[1]。以往单一方法检测三种性病病原体常会出现漏检。我们用PCR对本所性病门诊1995年4月～9月就诊500例泌尿生殖道感染患者同时进行了三种性病病原体联合检测,现报告如下。临床资料　共500例,男345例,女155例。年龄17～50岁,平均28.5岁。病程1周～3年。均有婚外性交史、配偶感染史或卖淫嫖娼史。依据非淋菌性尿道炎(NGU)和淋病的诊断标准[1]将病例分为两组。NGU组443例,淋病组57例。两组男女性别比分别为2.23∶1和2.16∶1。实验方法　标本取材和PCR具体操作方…     

连接酶链反应检测性病患者沙眼衣原体感染

目的 评价连接酶链反应(LCR)诊断性病患者尿道/宫颈中沙眼衣原体(Ct)的意义。方法 STD门诊尿道(宫颈)炎患者276例,取尿道/宫颈拭子,以LCR分析法检测Ct。采用每4份标本相混合的方法分别以LCR分析法检测尿道/宫颈拭子标本中的Ct,其中56例患者同时进行尿道/宫颈拭子Ct细胞培养。差异性结果由PCR法扩增Ct的主要外膜蛋白基因来进行确认,确定LCR分析法检测Ct的敏感性、特异性。结果 LCR分析法检测尿道/宫颈拭子Ct的敏感性、特异性分别为96.7%和100%。采用每4份标本相混合的方法分别以LCR分析法检测尿道/宫颈拭子标本中的Ct,与单独用每份标本逐一进行LCR检测比较,结果完全一致,符合率为100%。结论 以尿道/宫颈拭子为标本,LCR分析法检测Ct的敏感性、特异性高。适用于诊断泌尿生殖道Ct感染。用标本相混合的方法LCR分析检测尿道/宫颈拭子标本中的Ct,适用于Ct感染的普查。     

Comparison of the Amplicor Chlamydia trachomatis test and cell culture for the detection of urogenital chlamydial infections.

OBJECTIVE--To compare the polymerase chain reaction (PCR) Amplicor Chlamydia trachomatis test with the cell culture method, in diagnosing urogenital chlamydial infections. SUBJECTS--439 patients (327 women and 112 men) attending one STD clinic and Family Planning and Gynaecological Clinics in Lisbon, Portugal, between November 1993 and March 1994. METHODS--In women, two endocervical swab samples were collected: one for PCR Amplicor and one for standard culture technique. Men were asked to submit 20 ml of urine (first pass urine) for PCR Amplicor and one urethral specimen was taken for culture. The order of collection of the specimens was rotated every 50 patients. Discrepant results were further analysed by a second PCR with primers directed against the C trachomatis major outer membrane protein (MOMP) and by direct fluorescent antibody (DFA). RESULTS--After analysis of discrepancies, the adjusted sensitivity and specificity of PCR on endocervical specimens were 92.9% and 100% and the positive and negative predictive values were 100% and 99.7% respectively; on the urine samples these values were 100%, 99.1%, 100% and 99.1%, respectively. CONCLUSION--These results indicate that the PCR Amplicor test is a rapid sensitive and specific assay for the detection of C trachomatis in urogenital infections and provides a non-invasive technique for screening chlamydia infection in men.     

Detection of Chlamydia trachomatis in genital swabs: comparison of commercial and in house amplification methods with culture

AIMS: To evaluate the sensitivity of the Roche Cobas, Roche Amplicor plate kit, ligase chain reaction (LCR), and an in house polymerase chain reaction (PCR) by titration of purified elementary bodies (EB) and also to test 245 urethral and endocervical specimens for Chlamydia trachomatis by the four assays as well as conventional culture. STUDY DESIGN: EB titrations were run in duplicate in each commercial assay and six times in the in house PCR. Clinical samples were aliquoted and tested by each assay and were considered positive if C trachomatis was detected by two or more separate tests or if the sample was either culture or immunofluorescence positive. Major outer membrane protein (MOMP) specific primers were used as a confirmatory assay for the in house PCR. RESULTS: The in house PCR, Roche Cobas Amplicor, LCR, and Amplicor plate kit gave detection limits of approximately 1, 1-2, 2, and 2-4 EBs respectively. By the criteria described above for definition of a C trachomatis positive result in clinical samples we identified 23 true positives among the 245 clinical specimens. The in house PCR detected all 23 giving a sensitivity of 100% and a specificity of 98%. The Roche Cobas Amplicor, Roche Amplicor plate kit, and LCR detected 21, 19, and 19 of these respectively giving sensitivities of 87.5%, 82%, and 82% respectively and specificities of 99.5%, 99%, and 100% respectively. The culture gave a sensitivity of 78% and specificity of 100%. CONCLUSION: All four amplification assays had a greater sensitivity than the culture used routinely in this laboratory. The in house plasmid PCR had the greatest sensitivity and when combined with confirmation by immunofluorescence detected the greatest number of positives. This increased sensitivity is likely to have been achieved by the use of a DNA purification step and of nested primers in the amplification stage and their combined use in routine diagnostic assays for chlamydia might increase the frequency of C trachomatis detections. However, this assay is much less user friendly than the two semiautomated commercial assays investigated in this study.


     

Evaluation of patient-administered tampon specimens for Chlamydia trachomatis and Neisseria gonorrhoeae

BACKGROUND: The patient-administered tampon specimen has proven to be an easy and sensitive method for the diagnosis of genital Chlamydia trachomatis and Neisseria gonorrhoeae infections in women by polymerase chain reaction (PCR). This method avoids the need for endocervical sampling and stringent criteria for transport. GOAL: To evaluate two commercial amplification systems for the detection of C trachomatis and N gonorrhoeae from tampon specimens. STUDY DESIGN: A group of 400 positive and negative tampon specimens tested by an in-house PCR method were selected from a pool of more than 2,000 previously collected tampons. Overall, 93 C trachomatis-positive and 77 N gonorrhoeae-positive specimens were evaluated. Each specimen was tested by Roche Cobas Amplicor and Abbott LCx (LCR), and results were compared to the in-house PCR method. RESULTS: Detection of C trachomatis by both assays was not significantly different from the in-house PCR assay. Fewer tampons were positive for N gonorrhoeae by LCR than either the in-house assay (P = 0.0001) or by Roche Amplicor (P = 0.01). However, tampon specimens tested by Roche Amplicor required DNA extraction to achieve comparative sensitivity. CONCLUSION: Both commercial assays can be applied to tampon-collected specimens for automated detection of sexually transmitted diseases. The detection of C trachomatis was similar to the in-house PCR test for both assays (P = 0.73, 0.68). Detection of N gonorrhoeae resulted in fewer positive tampon specimens when tested by ligase chain reaction than both Roche Amplicor and in-house PCR.     

反向斑点杂交技术快速检测四种泌尿生殖道病原体

目的:建立一种能同时快速检测奈瑟淋球菌、沙眼衣原体、生殖支原体、单纯疱疹病毒1型和2型的杂交检测方法。方法:选取靶基因16S rRNA基因和gD糖蛋白基因,设计2对通用引物和特异性探针,建立双重PCR结合反向斑点杂交技术检测以上4种病原体,并对其检测能力进行评估。结果:建立的杂交检测方法检测奈瑟淋球菌、沙眼衣原体、生殖支原体、单纯疱疹病毒1型和2型的敏感性分别为1 CFU、3 IFU、6.7 pg/μL、1 CFU、1 CFU;各特异性探针只与相应病原体扩增产物杂交,未见交叉反应;与临床常规检测方法比较,一致率达96.8%,并显示能检出沙眼衣原体和生殖支原体双重感染。结论:该方法敏感性高、特异性好,并能同时检测多种病原体,在性病的快速诊断和病因学筛查等方面具有重要意义。     

Evaluation of Abbott Testpack Chlamydia for detection of Chlamydia trachomatis in patients attending sexually transmitted diseases clinics

This work compares a rapid solid-phase EIA (Abbott TestPack Chlamydia) to tissue culture and a direct fluorescent antibody test (Syva Microtrak) for detection of C. trachomatis in 436 patients attending two inner-city sexually transmitted diseases (STD) clinics. The prevalence of C. trachomatis by culture was 12% (5% in men, 15% in women). Overall sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of TestPack compared to culture were 70%, 98%, 80%, and 96% respectively. In men, 12 specimens were positive by TestPack, while only eight specimens were positive by culture. Six TestPack-positive, culture-negative specimens were further evaluated by centrifugation of culture transport media and examination of the sediment for chlamydia elementary bodies (EBs) using fluorescent monoclonal antibodies to C. trachomatis. Using this procedure, five of six culture negative specimens contained EBs (revised sensitivity 85%, specificity 99%, PPV 92%, NPV 99%). In 285 women evaluable in culture and TestPack, 44 (15%) specimens were culture positive; TestPack was positive in 29 (sensitivity 66%) culture positive women. Of 241 culture negative patients, 238 had negative TestPack results (specificity 99%) and no EBs were detected in the culture-negative, TestPack-positive specimens. Twenty-three (8%) Microtrak specimens were unsatisfactory for testing; two of these were culture and TestPack positive. Therefore, of 263 specimens evaluable using Microtrak, 42 (16%) specimens were culture positive; Microtrak was positive in 32 (sensitivity 76%) culture-positive women. Abbott TestPack Chlamydia is a rapid (25 minute), visually read format requiring no specialized equipment for detection of chalmydia infections with a sensitivity comparable to that of Microtrak.     

LCR技术检测男性首段尿中的淋病奈瑟菌和沙眼衣原体

目的：应用连接酶链反应（LCR）技术检测男性尿标本中的淋病奈瑟菌和沙眼衣原体,初步评价其敏感性和特异性。方法：采集受检者晨起或较长时间（2小时以上）不排尿后的首段尿（FVU）标本1131份,利用LCR技术对此尿液标本进行淋病奈瑟菌和沙眼衣原体检测,对Cut-off值在灰区以上的标本进行PCR检测。对LCR和PCR结果相异的标本,用另-LCR试剂进行复检,参照“扩大的金标准”来确定检测结果。结果：LCR技术检测淋病奈瑟菌的敏感性和特异性分别为100％和99．9％,沙眼衣原体分别为97．5％和98．4％。结论：应用LCR技术筛检男性尿液中淋病奈瑟菌和沙眼衣原体,是一种既敏感又特异的非侵入诊断方法,可避免取尿道标本给患者带来的痛苦。     

培养法及聚合酶链反应检测男性非淋菌性尿道炎患者中阴道毛滴虫

目的：研究男性非淋菌性尿道炎（NGU）患者中阴道毛滴虫感染的状况;评价尿液及尿道拭子聚合酶链反应（PCR）检测阴道毛滴虫感染的敏感性和特异性,建立一种非侵入性检测男性阴道毛滴虫感染的方法。方法：共收集105例患者。每例取2份尿道拭子标本,分别进行InPouchTV培养和PCR检测;取1份尿液标本进行PCR检测。以培养法为“金标准”,将两种PCR法与之比较,分别计算其敏感性、特异性、阳性预测值和阴性预测值。结果：InPouchTV培养法检出率为4．76％（5／105）;尿液PCR法检出率为3．81％（4／105）;尿道拭子PCR法检出率为4．76％（5／105）。尿液PCR法和尿道拭子PCR法的敏感性,特异性,阳性预测值和阴性预测值分别为80％．100％、100％、99％和80％、99％、80％、99％。结论：阴道毛滴虫感染是男性NGU的病因之一,在本研究中,阴道毛滴虫感染在男性NGU中的检出率为4．76％。尿液PCR法具有较高的敏感性和特异性且适用于非侵入性标本的检测,是具有临床应用价值的检测阴道毛滴虫的方法。