化学发光免疫法检测梅毒特异性抗体结果分析

目的 探讨化学发光法检测梅毒螺旋体特异性抗体的应用价值.方法 应用化学发光免疫测定法检测12503例住院患者梅毒螺旋体特异性抗体,并同ELISA、TPPA、RPR方法进行比较.结果 12503例住院患者化学发光法检出梅毒螺旋体特异性抗体阳性308例,同ELISA方法比较,阳性符合率99.08％,阴性符合率99.98％,总符合率99.93％.二者检测结果高度一致.同TPPA比较,阳性符合率98.7％.结论 化学发光免疫法检测梅毒特异性抗体具有特异性好、灵敏度高、结果易判断、便于自动化等优点,适合于梅毒的筛查.     

化学发光微粒子免疫分析在梅毒螺旋体抗体筛查中的应用探讨

目的 评价化学发光微粒子免疫分析法(CMIA)用于梅毒螺旋体抗体(TP-Ab)筛查的价值,并制定合理的TP-Ab血清学筛查方案.方法 收集在我院进行TP-Ab筛查的患者标本,共计30,100例,对于CMIA为阴性者直接报告结果,CMIA阳性者采用梅毒螺旋体明胶凝集试验(TPPA)确认,并同时进行快速血浆反应素环状卡片试验(RPR),分析结果的一致性.结果 在30100例筛查标本中,CMIA阳性者402例(1.3％),样本吸光度与临界质控吸光度比值(S/CO)在1.05～40.56之间.CMIA结果＞10 S/CO的标本共178例,TPPA阳性比例为100％,RPR阳性63例;在6-10 S/CO之间的标本共计50例,RPR阳性8例,TPPA阳性比例92％;在3～6 S/CO之间的标本59例,RPR阳性者5例,TPPA阳性的比例66.1％;在1～3 S/CO之间的标本115例,RPR均为阴性,TPPA阳性的比例仅为20％.结论 CMIA筛查TP-Ab具有较高的灵敏度,但特异性相对较低,采用CMIA进行TP-Ab筛查,并根据其S/CO值确定是否需要进一步采用另外一种高特异性梅毒螺旋体抗体检测试验进行确认,是TP-Ab筛查理想的方案,确认方法可以选择TPPA等试验.确认阳性者,应报告RPR结果,辅助临床区分既往感染和现症感染.     

梅毒螺旋体特异性抗体试验方法探讨及临床应用价值评估

目的 分析化学发光微粒子免疫法(CMIA)检测梅毒螺旋体特异性抗体的性能价值,探讨其在临床应用的意义.方法 回顾性分析北京市海淀医院2019年3月至2020年7月所有采用CMIA法进行梅毒螺旋体特异性抗体检测的标本61 051例,阳性标本采用梅毒螺旋体明胶颗粒凝集试验(TPPA)进行确证,分析其相关性.结果 61 0...     

免疫比浊法检测梅毒螺旋体抗体的方法评价

目的评价免疫比浊法在梅毒螺旋体抗体检测中的临床应用价值。方法分析免疫比浊法的精密度、线性、稳定性、干扰因素、携带污染指标,并与化学发光微粒子免疫分析法(CMIA)进行阳性率比较。结果免疫比浊法批内CV为1.64%~2.22%、批间CV为3.57%~3.74%;黄疸、脂血及溶血对本方法无明显干扰;阳性率均为16.8%,差异无统计学意义(x^2=0,P>0.05)。结论免疫比浊法检测结果与CMIA法检测结果相关性良好,可以满足临床诊断需要。     

老年患者梅毒螺旋体特异性抗体生物学假阳性结果分析

目的 分析我院老年患者梅毒螺旋体特异性抗体筛查情况,同时探讨其生物学假阳性的原因及临床意义.方法选取2016年1月至2016年12月我院8741例60岁以上老年患者,利用化学发光微粒子免疫检测法(CMIA)检测梅毒螺旋体特异性抗体,梅毒甲苯胺红不加热血清试验(TRUST)检测梅毒螺旋体非特异性抗体,同时采用免疫印迹法(Western Blot)进行梅毒确证.根据老年患者年龄将其分为60～69岁、70～79岁和80岁以上三个年龄组进行统计学分析.结果8741例60岁以上老年患者中有354例患者梅毒螺旋体特异性抗体阳性,60～69岁、70～79岁和80岁以上三个年龄组的梅毒螺旋体特异性抗体阳性率分别为3.42%、6.30%和6.02%,差异具有统计学意义(x2=31.236,P<0.001);同时,60~69岁与70~79岁年龄组比较,差异具有统计学意义(x2=28.932,P<0.001).240例老年患者CMIA筛查梅毒抗体阳性标本经TRUST和Western Blot检测都为阴性,且无临床症状和体征、无相关病史,结果判为假阳性.老年患者梅毒抗体假阳性人群主要分布在60~69岁.但随着老年患者年龄的增加,梅毒螺旋体抗体假阳性率也呈上升趋势,60~69岁、70~79岁和80岁以上三个年龄组的梅毒抗体假阳性率分别64.81%、71.43%和87.50%.老年患者梅毒螺旋体抗体生物学假阳性伴有的其他临床症状及指标异常主要包括癌症(55.36%)、溶栓剂或抗凝剂治疗(35.71%)、糖尿病(28.57%)、肝硬化(25.00%)、严重感染(25.00%)、肾病(17.86%)、自身抗体阳性(16.07%)、手术(14.29%)、肝炎(12.50%)和代谢紊乱(8.93%),各类原因相互影响,梅毒抗体检测假阳性S/CO值最高可达10.29.而梅毒螺旋体特异性抗体真阳性老年患者伴有的其他临床症状及指标异常百分比情况明显低于生物学假阳性患者.结论 对于CMIA梅毒筛查阳性而TRUST检测阴性的老年患者,需进行Western Blot检测并结合临床症状、体征和病史,作出最后诊断.老年患者随着年龄的增高,梅毒抗体的假阳性率也呈上升趋势,需注意癌症、自身免疫病、严重感染、代谢紊乱等可能会导致老年患者梅毒抗体检测生物学假阳性的因素,有助于临床医生分析检测结果并向患者作出合理解释.     

梅毒螺旋体抗体快速试剂的实验室性能评价

目的 使用梅毒螺旋抗体快速试剂国家参考品以及商品化的血清转化盘和不同临床时期的梅毒样本对目前市面主要使用的TP抗体快速检测试剂进行性能评价。方法 选取34批次梅毒螺旋抗体快速试剂对梅毒螺旋抗体快速试剂国家参考品以及商品化的血清转化盘和51份不同临床时期梅毒感染样本进行检测,分析不同试剂的检测性能差异并依据检测结果进行评分。结果 34批次梅毒螺旋抗体快速试剂有5批次产品不符合梅毒螺旋抗体快速试剂国家参考品的性能要求,均为阳性参考品符合率项目;34批次梅毒螺旋抗体快速试剂检测血清转换盘阳转天数最大相差4周时间;评价梅毒螺旋抗体快速试剂对51份临床样本检测阳性率为84.3%～100%。来自14个厂家的16批次试剂评分结果为84.3～99.4分。结论 部分梅毒螺旋抗体快速试剂对早期梅毒感染样本的检出率较低,一方面需要选取合适的评价样本,另一方面需要加强试剂生产的工艺稳定性。     

应用双抗原夹心ELISA法筛查献血员梅毒螺旋体抗体

目的：优选一种适宜于献血员梅毒筛查的试验方法。方法：采用检测梅毒特异性抗体的双抗原夹心ELISA法对献血员进行抗体检测，并与RPR检测结果进行比较，对两种方法检测结果不一致的标本，再用TPHA法进行确证。结果：ELISA法阳性检出率0．36％(41／1271)、RPR法阳性检出率0．26％(29／11271)。ELISA法与TPHA法总符合率97．5％(40／41)、RPR法与TPHA总符合率63．41％(26／41)。结论：ELISA法优于RPR法，具有较高的灵敏度和特异性，适宜于献血员的筛查．有利于控制和减少梅毒的输血传播。     

ELISA检测梅毒IgM抗体在早期梅毒诊断中的应用

为探讨ELISA法检测梅毒螺旋体IgM抗体（Tp-IgM)在早期梅毒诊断中的意义。本文对确诊的67例早期梅毒、3例早期神经梅毒,3例不定期潜伏梅毒患者与26名非梅毒对照。采用ELISA方法,进行血清/脑脊液的梅毒螺旋体IgM抗体检测,同时作RPR及TPPA检测,结果表明,ELISA,RPR及TPPA的特异性分别为100%、88.5%和100%;ELISA法对于早期梅毒的敏感性（95.5%)高于RPR和TPPA（均为83.6%),而在3例不定期潜伏梅毒血清和3例早期神经梅毒脑脊液中,ELISA各检出2例阳性,提示ELISA法检测Tp-IgM对早期梅毒的诊断价值优于RPR和TPPA,对早期神经梅毒和不定期潜伏梅毒的诊断不具优势。     

经化学发光免疫分析法检测梅毒抗体在梅毒患者早期诊断中的应用研究

目的:分析经化学发光免疫分析法(Chemiluminescence immunoassay,CLIA)检测梅毒抗体在梅毒患者诊断中的应用价值.方法:收集本院2019年1月至2021年2月收治且需行早期梅毒诊断的125例患者的临床资料,以梅毒螺旋体明胶凝集试验(Treponema pallidum gelatin agglutination,TPPA)检测结果作为金标准,比较甲苯胺红不加热血清学试验(Unheated serological test of toluidine red,TRUST)、CLIA检测诊断梅毒的敏感性、特异性、准确性及CLIA检测诊断效果.结果:125例患者中,TPPA检测出18例阳性,检出率为14.40％.TRUST检测出10例,检出率为8.00％.CLIA检测出21例,检出率为16.80,明显高于TRUST检测(P<0.05).但CLIA、TPPA检测比较无差异(P>0.05).TRUST检测对早期梅毒诊断敏感性、特异性及准确性分别为61.11％、98.13％、92.80％;CLIA检测分别为100.00％、97.20％、97.60％.CLIA检测诊断早期梅毒的敏感性显著高于TRUST检测(P<0.05).CLIA检测S/CO为1～5、6～10、>10者经TPPA检测为阳性的符合率分别为80.00％、100.00％、100.00％.结论:经CLIA检测梅毒抗体在梅毒患者诊断中具有较高的敏感性、特异性,且操作简便快捷,具有较高临床应用价值.     

Specific and sensitive diagnosis of syphilis using a real-time PCR for Treponema pallidum

A real-time PCR assay with a Taqman probe was developed that targeted the polA gene of Treponema pallidum. The test was validated using an analytical panel (n = 140) and a clinical panel of genital samples (n = 112) from patients attending a sexually transmitted infections clinic. High sensitivities and specificities of 94-100% were achieved using two real-time PCR platforms, the Rotor-Gene and the iCycler. The assay can be completed within 2 h, enabling reporting in <8 h. This fast and robust assay is suitable for implementation in routine laboratories for diagnosing primary syphilis.     

Evaluation of recomWell Treponema, a novel recombinant antigen-based enzyme-linked immunosorbent assay for the diagnosis of syphilis

Objective   To evaluate the diagnostic performance of an enzyme immunosorbent assay (recomWell Treponema) for the diagnosis of syphilis. The novel recombinant antigens Tpn47, TpN17 and TpN15 were utilized.
Methods   A total of 782 human serum specimens, belonging to four different categories (blood donors, n  = 200; routine laboratory screening for syphilis, n  = 400; syphilis patients, n  = 122; potential cross-reactors, n  = 60), were evaluated to compare the sensitivity and specificity of the recomWell Treponema kit with a standard whole Treponema pallidum cell lysate antigen-based ELISA (Syphilis Screening) and with micro-haemagglutination (MHA-TP).
Results   The overall specificity and sensitivity of the recomWell Treponema IgG was 98.9% and 98.3%, respectively. The specificity and sensitivity of Syphilis Screening ELISA was 98.7% and 98.3%, respectively. The agreement between recomWell Treponema and Syphilis Screening was 100%, 97.8%, 95.9% and 95% among the blood donor specimens, screening samples, syphilis specimens and the potential cross-reactors, respectively. Values of concordance varying from 96.7% to 98.3% were found in the different groups of sera between recomWell Treponema and MHA-TP. In addition, recomWell Treponema demonstrated a good diagnostic performance when used to detect the IgM to T. pallidum . No false-positive sera were identified and, in 17/19 samples from primary infection, an IgM immune response was found.
Conclusions   recomWell Treponema was shown to be a highly specific and sensitive method in all stages of syphilis screening and it can be considered as alternative to other ELISA tests based on native antigen preparations.     

Cell and Tissue Interactions of Treponema pallidum in Primary and Secondary Syphilitic Skin Lesions: An Ultrastructural Study of Serial Sections

There are limited reports on the ultrastructure of syphilis skin lesions. The aim of this study has been to perform an electron microscopic investigation of the morphology and the tissue distribution of treponemes in primary and secondary cutaneous lesions. Three cases of primary syphilitic chancre and one case of secondary syphilis were included. Prominent epidermal abnormalities in the primary chancre and a perivascular inflammatory infiltrate in the secondary lesion were found by light microscopy. Ultrastructurally, spirochetes were located mainly in the blood vessel walls and dermal tissue of the chancre lesions. In the secondary syphilis case, spirochetes were more abundant between epidermal keratinocytes. Most of them adjusted to the intercellular spaces. Occasionally, the electron microscopy images were highly suggestive of an intracellular location. Both the ultrastructural and immunohistochemical examination of the primary and secondary syphilis lesions showed a paradoxical distribution of the causative microorganisms compared to the light microscopic changes. In addition, the ultrastructural findings strongly suggest that Treponema pallidum subspecies pallidum invades tissues, not only through an intercellular, but also through a transcellular pathway.     

梅毒螺旋体抗原基因的克隆、表达及其在临床检验中的应用

目的　克隆、表达并纯化梅毒螺旋体相对分子质量为 47× 10 3 抗原 ,并检测梅毒患者的血清标本。方法　应用基因扩增技术分离此蛋白的全长基因 ,采用基因工程的方法 ,应用融合表达载体pGEX 2T ,重组、克隆得到带有梅毒螺旋体 47× 10 3 特异性抗原基因的重组菌株 ,经大肠杆菌表达系统获得重组融合蛋白。再经亲和层析获得纯品 ,并联合应用ELISA检测梅毒患者的血清标本 30份 ,同时检测非梅毒患者的血清标本 30份。结果　纯化后获得了梅毒螺旋体相对分子质量为 47× 10 3 的特异性抗原。ELISA检测梅毒患者血清结果均阳性。非梅毒患者血清均阴性 ,无非特异性交叉反应。结论　此项方法具有操作简便、准确的特点 ,为临床检测梅毒开辟了新的领域     

Magnitude and trend of HIV and Treponema pallidum infections among blood donors in Offinso-North District,Ghana: a nine-year retrospective,cross-sectional study

IntroductionBlood transfusion poses a high public health risk to recipients; hence no effort recommended to eradicate or minimize the danger of transmitting the infections.Reproductive Biology should be underestimated at minimizing the risk of TTIs. This study determined the prevalence and trend of HIV and syphilis infections in voluntary blood donors.MethodA retrospective analysis of secondary data from consecutive prospective voluntary blood donors who accessed Nkenkaasu District Hospital''s Blood Bank from January 2010 to December 2018 was conducted.ResultCumulatively, HIV and Treponema pallidum seropositivity identified in the present study was high (19.1%, [95% C.I (0.026–0.028)]). The prevalence of HIV and syphilis infections were 10.9% (95% C.I (0.098–0.120)) and 8.9% (95% C.I (0.073–0.92)) respectively. Prospective female blood donors were less likely to test positive for T. pallidum than males (OR 0.511, [0.340 – 0.769], p=0.001), but the infection was similar among different ages. The data showed downward trend for both HIV and T. pallidum seropositivity, (slope=-2.9467, p<0.0001) and (slope=-0.7117, p<0.0001) respectively.ConclusionSeroprevalence of HIV and Treponema pallidum were high, and their individual or combined seropositivity pose a significant threat to the safety of blood. Extensive and continuous screening for high-risk behaviours and infectious markers before blood donation is therefore Unit, Department of Obstetrics and Gynaecology, College of Medicine, Pan African University of Life and Earth Sciences Institute (PAULESI), University of Ibadan, Ibadan, Nigeria.     

Evaluation of an enzyme immunoassay technique for detection of antibodies against Treponema pallidum

In the present study, the performance of an enzyme-linked immunosorbent assay (ELISA) technique (Eti-syphilis-G and Eti-syphilis-M; DiaSorin) for detection of Treponema pallidum immunoglobulin M (IgM) and IgG antibodies for the laboratory diagnosis of syphilis was evaluated. Four hundred forty-one samples were studied. The sensitivity and specificity of the ELISA were 100 and 93%, respectively, compared with the results of a microhemagglutination assay for Treponema pallidum (MHA-TP) and 99.4 and 100%, respectively, compared with the results of the fluorescent treponemal antibody absorption (FTA-Abs) test. The results of the ELISA technique were concordant with those of MHA-TP for 98% of the samples tested, while the rate of concordance with the FTA-Abs test was 99.5%. The sensitivities of the rapid plasma reagin (RPR) test, MHA-TP, and the ELISA in the different phases of syphilis compared with the results of the FTA-Abs test were 92, 88, and 100%, respectively, for patients with primary syphilis; 100% for all tests evaluated for patients with secondary syphilis; 97.2, 99.4, and 100%, respectively, for patients with latent syphilis; and 57.9, 92.6, and 97.9%, respectively, for patients with past treated syphilis. The RPR test was reactive with 12 samples that were negative by all the specific tests. IgM antibodies were most frequently detected by the ELISA for IgM antibodies (32.8%) than by the FTA-Abs for IgM antibodies (28.4%). Detection of these antibodies by the FTA-Abs test and the ELISA for IgM antibodies decreased with the stage of disease (72 and 88%, respectively, for patients with primary syphilis to 17 and 19%, respectively, for patients with early latent syphilis). The high sensitivity and specificity of this ELISA technique during all stages of syphilis, together with the fact that it is a simple, objective, and easily automated method, lead us to believe that it could be used as a screening test for syphilis.     

ELISA法筛查临床样本中梅毒螺旋体抗体假阳性结果分析

目的:分析酶联免疫吸附试验(ELISA)从临床样本中检测出梅毒阳性结果中的假阳性及应对措施。方法:把5886例分为老年组和中青年组,用TP-ELISA从检出阳性91例,再用蛋白印迹法(WB)进行确认。结果:TP-ELISA法从老年组的梅毒抗体检出53例(2.64%),WB法确证45例,假阳性率15.09%;TP-ELISA法从中青年组的梅毒抗体检出38例(0.98%),WB法确证37例,假阳性率2.63%。且出假阳性时TP-ELISA法的S/CO值与真阳性时的S/CO值无显著性意义。结论:为提高老年人梅毒检测准确性,实验室使用WB法对有疑问的TP-ELISA法检测阳性样本进行确认是较好的选择。     

Evaluation of a new particle gel immunoassay for determination of antibodies against Treponema pallidum

A new particle gel immunoassay (DiaMed AG, Cressier sur Morat, Switzerland) with three recombinant Treponema pallidum antigens was evaluated with serum samples from patients with syphilis (n = 124) and patients without syphilis (n = 490). It proved to be a simple, rapid (20 min), and useful test with sensitivity, specificity, and positive and negative predictive values of 91.9, 99.8, 99.2, and 98%, respectively.