金标快速免疫试剂盒检测沙眼衣原体的性能评估

目的对目前最常用的两种胶体金标记快速免疫试验检测泌尿生殖道标本中沙眼衣原体的结果进行临床评估.方法共检测临床标本188例,包括男性尿道拭子标本52例,女性宫颈拭子标本136例.每例患者取拭子三支.前两支拭子标本分别用于两种金标快速试验("英国立明"及"美国思创").拭子使用顺序在每隔10例患者后作一交替.第三支拭子用于酶免法("Dako酶免")检测.结果以酶免法结果为参照,两种金标法对男性标本的敏感性分别为"英国立明"58.33%,"美国思创"为66.67%;对女性患者的敏感性分别为"英国立明"85.29%,"美国思创"82.35%.综合男女标本的检测结果,敏感性分别为"英国立明"78.26%,"美国思创"78.26%;特异性分别为92.96%、94.37%.结论两种金标类试剂检测结果之间差异无显著性,其敏感性均较低,尤其是对男性尿道拭子标本的漏检率较高.     

VIDAS CHL检测泌尿生殖道沙眼衣原体感染

目的探讨VIDAS CHL法用于性病患者尿道拭子标本以及男性首段尿(FCU)标本沙眼衣原体(CT)检测的可行性.方法使用组织培养法(TC)、VIDAS CHL检测法和聚合酶链反应(PCR)平行检测男性和女性拭子标本中CT,以组织培养为金标准,对VIDAS CHL和PCR进行评价.结果 232例男性标本TC阳性49例;VIDAS CHL和PCR法用于男性尿道拭子标本的敏感性分别为95.9%、93.9%,特异性分别为95.6%、94.5%,差异无显著性;VIDAS CHL法用于男性FCU的敏感性和特异性分别为85.7%和96.7%,与拭子标本作自身配对比较,检测结果无差异.151例女性拭子标本,TC法阳性23例,VIDAS CHL和PCR法的敏感性分别为100%和95.7%,特异性分别为96.1%、93.8%,差异无显著性.结论 VIDAS CHL法用于性病患者男性和女性拭子标本的CT检测,具有很高的敏感性和特异性,用于男性患者FCU的CT检测也是可行的;对于CT阳性率较高的性病人群,VIDAS CHL可以不做阻抑证实试验.     

VIDAS CHL检测泌尿生殖道沙眼衣原体感染

目的 探讨VIDAS CHL法用于性病患者尿道拭子标本以及男性首段尿（FCU）标本沙眼衣原体（CT)检测的可行性。方法 使用组织培养法（TC）、VIDAS CHL检测法和聚合酶链反应（PCR）平行检测男性和女性拭子标本中CT，以组织培养为金标准，对VIDAS CHL和PCR进行评价。结果 232例男性标本TC阳性49例；VIDAS CHL和PCR法用于男性尿道拭子标本的敏感性分别为95.9%、93.9%，特异性分别为95.6%、94．5％，差异无显著性；VIDAS CHL法用于男性FCU的敏感性和特异性分别为85．7％和96．7％，与拭子标本作自身配对比较，检测结果无差异。151例女性拭子标本，TC法阳性23例，VIDAS CHL和PCR法的敏感性分别为100％和95．7％，特异性分别为96.1%、93.8%，差异无显著性。结论 VIDAS CHL法用于性病患者男性和女性拭子标本的CT检测，具有很高的敏感性和特异性，用于男性患者FCU的CT检测也是可行的；对于CT阳性率较高的性病人群，VIDAS CHL可以不做阻抑证实试验。     

金标法与酶联免疫法(ELISA)在筛选HIV患者的应用效果比较

分别采用金标法和酶联免疫法对1000份待测样本进行HIV抗体检测,从中选出呈现阳性的标本;用同一种方法对检测出的阳性标本进行复查;重新抽取阳性标本患者血液,送往艾滋病确诊实验室进行检测。结果 1000份样本中,金标法与酶联免疫法初次筛查出的阳性标本数分别为11与8份,经实验室确诊的阳性标本数为7份;金标法与酶联免疫法的假阳性率分别为36.4%和12.5%。金标法虽敏感性较高但假阳性率较高,酶联免疫法敏感性较为理想且综合准确率高于金标法,二者分别适合于检测数量大小不同的样本。     

连接酶链反应和细胞培养检测男性尿液中沙眼衣原体

目的　评价连接酶链反应(LCR)检测男性尿液标本中沙眼衣原体(Ct)的敏感性和特异性。方法　取 852例患者尿道拭子作Ct培养;同时取患者较长时间(2h以上)不排尿后的首次尿 (FVU)标本,用质粒LCR检测Ct。对培养法和质粒LCR检测结果相异的标本,用另一针对Ct主要外膜蛋白基因的引物进行LCR确证,参照“扩大的金标准”确定检测结果。结果　质粒LCR敏感性和特异性分别为 98. 6%和 99. 4%,而培养法分别为 77. 4%和 99. 5%,前者敏感性高于后者 (P<0. 001)。有、无尿道症状的患者间LCR敏感性和特异性无显著性差异。结论　LCR是一种既敏感又特异的Ct感染诊断方法。     

无痛型尿道拭子在男性患者尿道分泌物取样中的应用

选取30例男性不孕且需经尿道分泌物取样的患者作为观察对象。所有入选对象均采用无痛型尿道拭子和传统尿道拭子分别进行取样。将两种方式取得的样本均进行解脲支原体的检测,并询问患者取样时的疼痛情况。结果无痛型尿道拭子组无疼痛、轻度疼痛、中度疼痛、重度疼痛的发生率依次分别为36.67%、56.67%、6.66%、0%,传统尿道拭子组依次分别为0、20%、36.67%、43.33%,两组之间存在明显差异(P<0.01);无痛型尿道拭子组解脲支原体阳性检出率为33.33%,传统尿道拭子组为40%,两组之间无差别(P>0.05)。在男性患者尿道分泌物取样中应用无痛型尿道拭子可有效的减轻疼痛,且能达到与传统尿道拭子相同的检测效果。     

梅毒螺旋体两种检测方法的比较

目的为了探讨金标法检测梅毒螺旋体的敏感性。方法采用酶联免疫吸附试验(ELISA)法对本院1189例临床样本进行初筛试验,再对初筛结果阳性的30例血清进行金标法梅毒螺旋体检验。结果酶联免疫吸附试验(ELISA)阳性的30例样本中,金标法阳性28例,二者符合率为93.3%。结论ELISA法和金标法在敏感性方面有较高的符合率,适合于标本量较少的基层医院和急诊检验。     

2种免疫层析法试剂检测沙眼衣原体比较

目的比较2种免疫层析法(ICA)试剂盒检测沙眼衣原体(CT)符合率.方法临床送检的228例标本分别用2种进口ICA试剂盒进行检测.结果测试标本中,美国Quidel公司生产的Quick Vue(QV)试剂盒与英国Unipath公司生产的Clearviw Chlamydia(CC)试剂盒检测CT,总符合率为98.68%.其中男性尿道拭子标本的符合率为99.36%,女性宫颈标本的符合率为97.22%.结论 QV与CC对检测泌尿生殖道Ct感染有较高的符合率.     

2种免疫层析法试剂检测沙眼衣原体比较

目的　比较 2种免疫层析法 (ICA)试剂盒检测沙眼衣原体 (CT)符合率。方法　临床送检的 2 2 8例标本分别用 2种进口ICA试剂盒进行检测。结果　测试标本中 ,美国Quidel公司生产的QuickVue(QV)试剂盒与英国Unipath公司生产的ClearviwChlamydia(CC)试剂盒检测CT ,总符合率为 98.6 8%。其中男性尿道拭子标本的符合率为 99.36 % ,女性宫颈标本的符合率为 97.2 2 %。结论　QV与CC对检测泌尿生殖道Ct感染有较高的符合率。     

全自动荧光酶免疫分析仪检测泌尿生殖道衣原体的评价

目的：评价生物梅里埃公司的全自动荧光酶免疫分析系统（VIDAS）检测衣原体（CHL）的敏感性和特异性。方法：应用VIDAS检测CHL与细胞培养比较,不符者使用直接荧光抗体技术检验证;同时使用CHL菌株D和E血清型,应用上述两种方法,进行敏感性的比较,结果：163份性病科门诊男女患者尿道和宫颈拭子标本中,细胞培养阳性者33份（20．2％）以扩大的金标准计算,其敏感性为80．5％,特异性是1005。VIDAS检测出CHL44份（27．05）阳性标本,敏感性和特异性分别为95．3％和97．65。CHL菌株D和E血清型敏感性中,培养阳性的最高滴度分别为1：102400和1：51200,VIDAS检测CHL的阳性的最高滴度均为1：6553600。结论：与扩大的金标准比较,VIDAS检测CHL具有良好的敏感性和特异性。该仪器操作简便,检测时间仅为1h。此外,还可用于尿液标本中的沙眼衣原体检测,避免了男性尿道拭子标本采集的痛苦。     

Diagnosis of Neisseria gonorrhoeae and Chiamydia trachomatis infection using first-voided urine in men with urethritis

A study was designed to examine the feasibility of making the specific diagnoses of gonorrhoea and chlamydia infection from a single specimen of urine. Urine specimens and urethral swabs were collected from 212 male attenders at a genitourinary medicine clinic who had evidence of urethritis. Urethral swabs and urine sediments were cultured for Neisseria gonorrhoeae and urine sediments were also tested by Gonozyme and IDEIA enzyme immunoassays for N. gonorrhoeae and Chlamydia trachomatis antigens respectively Sixty-three urethral cultures were positive for N. gonorrhoeae and 83 tests were positive in the routine amplified immunoassay for chlamydia antigen. Nineteen patients had a dual infection of gonorrhoea and chlamydia. The incidence of chlamydia in non-gonococcal urethritis was 43%, this closely agreed with our previous studies. Urine deposit and urethral swab culture for N. gonorrhoeae gave a concordant negative result in 148 patients and urine culture detected 61 out of 63 patients found positive by urethral swab culture, a sensitivity of 96.8%. The immunoassay for gonococcal antigen in urine detected 60 out of 63 urethral culture positive patients. If two persistently equivocal results were taken as reactive then the sensitivity of the test was 98.4% with a specificity of 94%. Our results showed that both N. gonorrhoeae and C. trachomatis can be detected readily using appropriate enzyme immunoassays on a single urine sample from symptomatic males. This approach to sexually-transmitted disease (STD) screening may be applicable to mass populations.     

A real-time PCR assay for the detection of Neisseria gonorrhoeae in genital and extragenital specimens

A Neisseria gonorrhoeae LightCycler (NGpapLC) assay targeting the porA pseudogene was compared with bacterial culture for detection of N. gonorrhoeae in 636 clinical specimens (216 cervical, 185 urethral, 196 throat, and 39 rectal swab specimens). The specificity of the NGpapLC assay was further investigated by testing a bacterial reference panel comprising several Neisseria species. Overall, 19 (3.0%) specimens were positive and 613 (96.4%) specimens were negative by both methods. Four (0.6%) specimens were positive by the NGpapLC assay only. For the cervical and urethral swabs, the NGpapLC provided 100% sensitivity and 100% specificity compared with bacterial culture. Following discrepant analysis, the clinical sensitivity and specificity of the NGpapLC for throat and rectal swabs was also 100%. For the bacterial panel, only N. gonorrhoeae isolates provided positive results. The results show the NGpapLC assay is suitable for use on a range of clinical specimens and could improve detection of pharyngeal N. gonorrhoeae.     

Diagnosis of Chlamydia trachomatis and Neisseria gonorrhoeae. Genitourinary infections in males by the Amplicor PCR assay of urine

The Amplicor CT/NG polymerase chain reaction (PCR) test on urine specimens from males was prospectively evaluated against established specimens and laboratory methods for diagnosing Chlamydia trachomatis and Neisseria gonorrhoeae genitourinary infections, in patients from a remote region of Western Australia. Seventy-three males who were tested for both C. trachomatis and N. gonorrhoeae by both conventional methodology and Amplicor PCR on urine were enrolled in the study. Established testing comprised enzyme immunoassay/immunofluorescence antigen testing (EIA/IF) for C. trachomatis and microscopy and/or culture for N. gonorrhoeae on urethral swabs. Positive test results were confirmed using a set of criteria that included supplemental PCR testing and clinical history. Overall, 13.7% of patients were resolved as positive for C. trachomatis and 52.1% as positive for N. gonorrhoeae. The sensitivity and specificity of the Amplicor CT/NG PCR on male urine specimens for C. trachomatis were 80.0% (8/10) and 95.2% (60/63), compared with 60.0% (6/10) and 100.0% (63/63) for EIA/IF on urethral swabs. For N. gonorrhoeae, the sensitivity and specificity of the Amplicor CT/NG PCR on male urine specimens were both 100% (38/38 and 35/35, respectively) compared with 86.8% (33/38) and 100% (35/35) for microscopy and/or culture on urethral swabs. The results of this study indicate that the Amplicor CT/NG multiplex PCR test for C. trachomatis and N. gonorrhoeae performed on urine in males provides a highly sensitive, specific, and robust method for the diagnosis of both C. trachomatis and N. gonorrhoeae, for the early detection of both symptomatic and asymptomatic infected individuals.     

Evaluation of Seeplex® STD6 ACE Detection kit for the diagnosis of six bacterial sexually transmitted infections

Traditionally, the diagnosis of bacterial sexually transmitted infection (STI) has been dependent on the isolation of the causative pathogens by culturing endocervical or urethral swab specimens on selective media. While such procedures typically provide excellent diagnostic accuracy, they are often time-consuming and expensive. A multiplex polymerase chain reaction (PCR) assay, based on a semi-automated detection system, was evaluated for the detection of six STI causative organisms. The Seeplex^® STD6 ACE (auto-capillary electrophoresis) Detection assay employed six pairs of dual priming oligonucleotide (DPO?) primers specifically targeted to unique genes of Chlamydia trachomatis, Neisseria gonorrhoeae, Mycoplasma genitalium, Ureaplasma urealyticum, Mycoplasma hominis, and Trichomonas vaginalis. A total of 739 specimens (304 cervical swabs and 435 urine samples) collected for 4 months were tested, and results were compared to those obtained with a combined monoplex PCR. The concordance between the multiplex PCR and monoplex PCR assay was 100% for both sensitivity and specificity. We also tested for the presence of two pathogenic bacteria (C. trachomatis and N. gonorrhoeae) and compared the results obtained with the multiplex PCR and BD ProbeTec duplex strand displacement amplification (SDA). The results of the multiplex PCR and duplex SDA were 99.7% concordant for C. trachomatis and 100% concordant for N. gonorrhoeae. The multiplex PCR assay using the Seeplex^® STD6 ACE Detection kit proved to be a novel cost-effective and fast diagnostic tool with high sensitivity and specificity for the simultaneous detection of six STI pathogens.     

Detection of Chlamydia trachomatis in symptomatic and asymptomatic populations with urogenital specimens by AMP CT (Gen-probe incorporated) compared to others commercially available amplification assays

The present study was designed to evaluate the sensitivity and specificity of AMP CT (Gen-Probe Incorporated, San Diego, CA, USA) on urogenital specimens taken from symptomatic patients and on first void urine (FVU) specimens from asymptomatic patients. In symptomatic patients, 618 specimens from 140 men (140 urethral swabs and 140 FVU) and 202 women (202 endocervical swabs and 136 FVU) were tested by using cell culture, AMP CT and Amplicor Chlamydia trachomatis MWP (microwell plate) (Roche Diagnostics, Somerville, NJ, USA) on genital samples, and AMP CT and Amplicor on FVU. A clinical specimen was considered to be truly positive if either the cell culture was positive and/or both AMP CT and Amplicor were positive. In the asymptomatic population, a total of 300 FVU (136 women and 164 men) were tested by four amplification methods, AMP CT, LCx (Abbott, Abbott Park, IL, USA), Amplicor MWP, and Cobas Amplicor. A subject was considered to be infected when two or more amplification methods were positive. In the symptomatic population (prevalence 13%), concordant results were observed in 320/342 cases (93.5%). After analysis of discordant results, the sensitivity of AMP CT, Amplicor, and culture was 100%, 95.5%, 68.8%, respectively, and the specificity was 98.3%, 99.3%, 100% respectively. The number of false negative results by AMP CT in urine, probably due to labile inhibitors, was 3/276 (1%). In the asymptomatic population, the results were concordant in 298/300 (99.3%), seven positive and 291 negative. Two results were considered false positives, one by Cobas Amplicor, one by AMP CT. Compared to other amplification methods, AMP CT is at least as sensitive for the identification of chlamydial infection in symptomatic and asymptomatic men and women on genital or urine specimens.     

Detection ofUreaplasma urealyticum in urethral swab samples from asymptomatic men and men with urethritis by a polymerase chain reaction-based assay

We compared a polymerase chain reaction (PCR)-based assay with primers specific for the 16S rRNA gene ofUreaplasma urealyticum with culture techniques for detectingU. urealyticum from urethral swab samples obtained from 256 asymptomatic men. Of 24 samples positive forU. urealyticum by culture, 23 samples were positive by the PCR-based assay, whereas 2 of 232 samples with a negative culture were positive by the PCR-based assay. The sensitivity and specificity for the PCR-based assay compared to culture were 95.8% and 99.1%, respectively. Our results confirmed the validity of the PCR-based assay for identifying this pathogen from urethral swab samples. In this study, we also examined urethral swab samples obtained from 195 men with urethritis for the detection ofU. urealyticum by this assay.U. urealyticum was detected in 1 (3.4%) of 29 men with gonococcal urethritis and 23 (13.9%) of 166 men with nongonococcal urethritis. This assay may be a relevant tool to diagnose urethralUreaplasma infections.     

Use of the polymerase chain reaction for the detection of Chlamydia trachomatis from endocervical and urine specimens in an asymptomatic low-prevalence population of women

The Amplicor Chlamydia trachomatis test is a polymerase chain reaction (PCR)-based methodology used for the detection of a cryptic plasmid found in C. trachomatis. It was evaluated in comparison with cell culture and the Microtrak II Chlamydia enzyme immunoassay (EIA) for the detection of C. trachomatis in urogenital specimens from women. Endocervical swabs were collected from 993 women attending the women's unit at the Mount Sinai Hospital in Toronto. In addition, concomitant first void urine specimens were collected from 394 of these women for PCR testing only. As compared with culture of the endocervical specimens, PCR and EIA had a sensitivity, specificity, positive predictive value and negative predictive value of 84.6%, 99.2%, 57.9%, and 99.8% and 61.5%, 99.7%, 72.7%, and 99.5%, respectively. As compared with the secondary gold standard of a positive culture and/or a positive PCR using a primer to the major outer membrane protein the sensitivity, specificity, positive, and negative predictive values for culture were 72.2%, 100%, 100%, and 99.5%, respectively. For the Amplicor PCR and EIA the results were 88.9%, 99.7%, 84.2%, and 99.9% and 61.1%, 99.9%, 91.7%, and 99.6%, respectively. When the urine PCR was compared with the same standard, the test had a sensitivity of 91.7% and a specificity of 99.5%. Based on this study the Amplicor C. trachomatis test was found to be sensitive and specific for the detection of C. trachomatis in both endocervical and urine specimens.