梅毒胶体金法在临床诊断中的应用价值

目的了解梅毒胶体金法在自愿咨询检测(VCT)门诊、社区卫生服务中心和非政府组织(NGO)工作室等临床诊断中的应用价值。方法应用梅毒螺旋体(TP)抗体检测试剂(胶体金法)、酶联免疫吸附试验(ELISA)、梅毒螺旋体明胶颗粒凝集试验(TPPA),同时对2011-2013年医院哨点监测性病门诊送检的血清标本进行检测。结果对1204例样本进行检测,梅毒胶体金法阳性检出率为20.51%(247/1204),ELISA法为21.01%(253/1204),TPPA法为20.02%(241/1204)。以TPPA为金标准,梅毒胶体金、ELISA的敏感性均为99.59%,特异性分别为99.27%、98.65%。胶体金法与TPPA法和ELISA法经Kappa一致性检验比较(u分别为135.20、137.71,k=0.98,P0.05),具有极强的一致性。结论梅毒胶体金检测敏感性高、特异性强,具有简便、迅速的特点,值得在临床诊断中推广应用。     

四川省2021年梅毒血清学试验室间质量评价结果分析

<正>梅毒血清学实验室检测包含两类血清学试验,一类是梅毒螺旋体血清学试验,包括：TPPA、梅毒螺旋体血凝试验(treponema pallidum hemagglutinationassay,TPHA)、ELISA、快速检测试验(rapid test,RT)、化学发光免疫试验(chemiluminescence immunoassay,CLIA)等;另一类是非梅毒螺旋体血清学试验,常用的有RPR、TRUST^[1]。     

CLIA法联合TPPA法在临床梅毒血清学筛查中的应用评价

目的探讨化学发光免疫分析(CLIA)法联合梅毒螺旋体明胶颗粒凝集试验(TPPA),在临床梅毒血清学筛查中的应用价值。方法用CLIA进行特异性梅毒螺旋体抗体初筛试验,阳性标本再经TPPA和梅毒甲苯胺红不加热血清试验(TRUST)复检。CLIA与TPPA结果不一致者用蛋白印迹法(WB)验证。结果 599例CLIA初筛阳性标本用TPPA法复检阳性568例、阴性31例;TRUST法复检阳性189例、阴性410例。31例TPPA阴性标本经WB试验验证结果为18例阴性、13例阳性。结论 CLIA法联合TPPA法初筛梅毒作为梅毒筛查的流程是可行的,值得推广。     

酶联免疫吸附试验检测梅毒患者血清中抗苍白螺旋体IgM抗体

目的评价抗苍白螺旋体IgM抗体检测对梅毒的临床意义.方法用酶联免疫吸附试验(ELISA)对72例梅毒患者检测了特异性IgM抗体,并与快速血浆反应素环状卡片试验(RPR)、梅毒螺旋体明胶颗粒凝集试验(TPPA)的检测结果进行比较分析.结果血清抗苍白螺旋体IgM抗体在一期梅毒的阳性率为73.3%(11/15),在二期梅毒的阳性率为88.9%(16/18),二者差异无显著的统计学意义(χ2＝1.6363,P＞0.10).在潜伏梅毒,IgM抗体阳性率为26.1%(6/23),非常显著地低于早期显性梅毒(χ2＝17.6189,P＜0.005).在一期、二期和潜伏梅毒,RPR和TPPA的阳性率均为100%.入组前2～24个月已经正规抗梅治疗的梅毒16例,其中IgM抗体阳性2例.结论 ELISA法检测特异性IgM抗体诊断一期梅毒并不优于RPR和TPPA.IgM抗体在潜伏梅毒敏感性低,其诊断应依靠RPR和TPPA.目前不推荐单独检测抗梅毒IgM抗体来监测病情和判断疗效.抗苍白螺旋体IgM抗体的临床意义有待更深入的研究.     

三种梅毒血清学检测策略在临床梅毒诊断中的应用研究

目的探讨不同梅毒血清学检测策略在梅毒诊断中的应用。方法按照正向策略、反向策略和欧洲疾病控制与预防中心(ECDC)反向策略三种流程,采用甲苯胺红不加热血清学实验(TRUST),梅毒螺旋体抗体明胶颗粒凝集试验(TPPA)和梅毒螺旋体抗体化学发光试验(CLIA)检测所有血清标本,比较3种检测策略对梅毒的检出价值。结果选择865例梅毒患者作为梅毒组,再选择同期排除梅毒的患者100例作为对照组,3种方法对不同分组的梅毒检出率差异均有统计学意义(P均0.05);TRUST、CLIA和TPPA的检出敏感性分别为87.28%,99.88%,99.54%,特异性分别为98.00%,97.00%,100.00%,准确度分别为88.39%,99.59%,99.59%;正向策略、反向策略和ECDC反向策略的敏感性分别为85.20%、99.42%、99.54%,特异性分别为98.00%、97.00%、100.00%,准确度分别为86.53%、99.17%、99.59%。结论不同梅毒血清学检测方法检测性能不一致,反向策略或ECDC反向策略具有更好的敏感性、特异性和准确度,适用于临床筛查和诊断。     

化学发光微粒子免疫分析法检测孕妇梅毒特异性抗体的应用评价

目的 探讨化学发光微粒子免疫分析法(CMIA)检测孕妇血清中梅毒螺旋体特异性抗体(TP-Ab)的应用价值.方法 150 314例妊娠期妇女血清标本采用CMIA检测TP-Ab,S/CO≥0.5用TPPA及ELISA检测;以TPPA为确证方法,比较不同S/CO值CMIA与TPPA的符合率.CMIA与TPPA检测结果不一致的...     

两种梅毒血清学检测方法的联合运用分析

目的进一步提高酶联免疫吸附试验(ELISA)对梅毒的筛查率,确保梅毒血清学的检测效果。方法采用ELISA联合甲苯胺红不加热血清试验(TRUST),对2011年上海浦东新区2 686例监测人群的血清样本进行梅毒血清学筛查,联合检测的血清学阳性样本用梅毒螺旋体明胶凝集试验(TPPA)甄别其生物学假阳性;再从2 397份两种方法均为阴性的样本中,随机抽取1 000份进行TPPA检测,甄别其生物学假阴性。结果 ELISA法相对于TPPA法的敏感性为98.9%,特异性为98.4%,两法间差异无统计学意义(P>0.05);ELISA与TRUST法联合应用后的敏感性接近100%,特异性为98.3%。结论 ELISA与TRUST方法的联合应用,其敏感性接近100%,表明其可用于梅毒筛查;另一方面,两者联合运用的特异性为98.3%,具有一定的假阳性率,表明其阳性样本(两法均为阳性的样本除外)还需应用TPPA方法进行最终确认。     

2种化学发光仪检测梅毒螺旋体特异性抗体性能分析

目的:以梅毒螺旋体免疫印迹法(TP-WB)为金标准,比较科美和雅培2种化学发光仪在检测梅毒特异性抗体时的差异。方法:选取科美检测梅毒阳性样本80例和雅培检测梅毒阳性样本80例,160例样本分别用2种化学发光检测梅毒特异性抗体后再用WB方法复检。结果:科美CLIA阳性预测值为90.07%,阴性预测值为88.89%,符合率为90.00%。雅培CMIA阳性预测值为93.19%,阴性预测值为100.00%,符合率为93.75%,两者差异无统计学意义(χ~2=0.950,P=0.330;χ~2=1.513,P=0.219;χ~2=1.507,P=0.220)。当科美S/CO9.01时阳性预测值为100%,雅培S/CO6.01时阳性预测值为100%,含有P47特征条带的样本数为108例(78.83%),为所有特征条带中出现频率最高。结论:科美和雅培化学发光在检测梅毒时与WB法比较均有较高符合率,雅培S/CO值≥6.01、科美S/CO值≥9.01时检测结果即为真阳性S/CO值,临床工作中可以使用价格较低廉的科美CLIA作为雅培CMIA的有效补充。     

2种化学发光仪检测梅毒螺旋体特异性抗体性能分析

目的:以梅毒螺旋体免疫印迹法(TP-WB)为金标准,比较科美和雅培2种化学发光仪在检测梅毒特异性抗体时的差异。方法:选取科美检测梅毒阳性样本80例和雅培检测梅毒阳性样本80例,160例样本分别用2种化学发光检测梅毒特异性抗体后再用WB方法复检。结果:科美CLIA阳性预测值为90.07%,阴性预测值为88.89%,符合率为90.00%。雅培CMIA阳性预测值为93.19%,阴性预测值为100.00%,符合率为93.75%,两者差异无统计学意义(χ~2=0.950,P=0.330;χ~2=1.513,P=0.219;χ~2=1.507,P=0.220)。当科美S/CO9.01时阳性预测值为100%,雅培S/CO6.01时阳性预测值为100%,含有P47特征条带的样本数为108例(78.83%),为所有特征条带中出现频率最高。结论:科美和雅培化学发光在检测梅毒时与WB法比较均有较高符合率,雅培S/CO值≥6.01、科美S/CO值≥9.01时检测结果即为真阳性S/CO值,临床工作中可以使用价格较低廉的科美CLIA作为雅培CMIA的有效补充。     

两种CLIA系统检测梅毒抗体的一致性及临界值设置

目的观察不同化学发光(CLIA)系统检测梅毒(TP)抗体与梅毒螺旋体明胶颗粒凝集试验(TPPA)的一致性并设定临界值。方法以博阳LICA500检测抗TP抗体S/CO1.0的血清样本296例﹑S/CO1.0的血清样本50例;同时以雅培i2000SR复检,以TPPA法进行确认检验。以TPPA为标准,方法间一致性比较采用Kappa检验;以TPPA为标准,绘制抗TP抗体S/CO比值受试者工作特征曲线(ROC曲线),得到约登指数最大时的S/CO比值以设定其临界值。结果雅培i2000SR抗TP抗体与TPPA检测结果的一致性较好(Kappa=0.826),博阳LICA500抗TP抗体与TPPA检测结果的一致性较差(Kappa=0.438);两者抗TP抗体与TPPA结果间的差异均具有显著性(P0.05)。雅培i2000SR检测抗TP抗体的灵敏度为100%,特异度为82.8%,阳性预测值为88.9%,阴性预测值为100%;而LICA500检测抗TP抗体的灵敏度为100%,特异度为61.9%,阳性预测值为73.0%,阴性预测值亦为100%。ROC曲线分析显示,博阳LICA500和雅培i2000SR检测抗TP抗体的曲线下面积分别为0.986和0.989;博阳LICA500抗TP抗体最佳诊断界值为10.095,其灵敏度为96.8%,特异度为97.5%;雅培i2000SR抗TP抗体最佳诊断界值为3.02,其灵敏度为95.8%,特异度为100%。结论不同化学发光系统检测梅毒抗体的诊断效能有差异,各实验室应设置最佳诊断界值并验证后使用。     

三种实验室检测方法对梅毒诊断的临床意义分析

目的检验三种梅毒实验室检测方法的梅毒阳性检出率,分析各检测指标间的相互关系,探讨其临床意义。方法抽取性病门诊就诊者的静脉血,用梅毒酶联免疫吸附试验(ELISA)检测梅毒抗体,同时用梅毒螺旋体明胶凝集试验(TPPA)检测同一份血清,二种方法均为阳性者用荧光梅毒螺旋体抗体吸收法(FTA-ABS)检测及免疫印迹法(WB)确认,对试验数据进行统计分析和检查。结果在所有784名性病门诊就诊者中,ELISA检测梅毒抗体阳性者306人,阳性率39.03%;TPPA法检测梅毒抗体阳性为297人,阳性率为37.88%;经FTA-ABS检测确证阳性299人,阳性率为38.14%;最终WB结果与FTA-ABS检测一致。结论为了提高梅毒的检出率及正确性,对高度疑似梅毒的患者,应同时进行两种抗体的检查。     

The sensitivity of syphilis assays in detecting different stages of early syphilis

Our aim was to determine the sensitivity of the Murex ICE enzyme immunoassay (EIA) as a screening test for early syphilis and to determine how many additional cases of infection were detected by performing additional tests when requested on clinical grounds.This was an observational study on consecutive patients diagnosed with syphilis in the Department of Genitourinary Medicine, Edinburgh between January 1st 2004 and April 1st 2005. Additional tests were performed on sera that gave a positive or equivocal EIA on screening as well as by clinical request on sera from contacts of syphilis, and those with clinical signs of syphilis. Additional tests included a Venereal Diseases Research Laboratory (VDRL) carbon antigen test, a Treponema pallidum particle agglutination (TPPA) test, INNO-LIA line immunoblot assay, and an EIA specific for anti-treponemal IgM.A total of 105 patients were diagnosed with syphilis: primary (50), secondary (26), early latent (8), and of unknown duration (21). The TPPA was the most sensitive test in primary syphilis and had a sensitivity of 96% (48/50), which was significantly higher (P <0.05) than the sensitivity of 84% (42/50) for the screening EIA: seven of the EIA negatives were detected by EIA-IgM, six by TPPA, five by immunoblot, and two by VDRL. EIA-IgM was negative in six primaries; all were positive by TPPA and immunoblot.We conclude that, in order to maximize the serological detection of primary syphilis a specific EIA-IgM test and a TPPA test should be performed whenever there is a clinical suspicion of primary infection. This is particularly important when an EIA such as Murex ICE is used as a single screening test as it is less sensitive than the TPPA in primary infection.     

Laboratory diagnosis of heparin‐induced thrombocytopenia

Heparin‐induced thrombocytopenia (HIT) is a clinical‐pathological disorder; thus, laboratory testing for the pathogenic platelet‐activating antiplatelet factor 4 (PF4)/heparin antibodies is central for diagnosis. The “iceberg” model summarizes the inter‐relationship between platelet activation assays and PF4‐dependent immunoassays, with platelet‐activating antibodies comprising a subset of anti‐PF4/heparin antibodies. The platelet serotonin‐release assay (SRA), performed by reference laboratories, has high sensitivity and specificity for HIT (~95% each), and is especially suited for detecting highly pathogenic HIT sera containing both heparin‐dependent and heparin‐independent platelet‐activating antibodies; this latter subgroup of antibodies explains “autoimmune HIT” disorders (delayed‐onset, persisting, spontaneous, heparin “flush,” fondaparinux‐associated). Recently, SRA‐negative HIT has become recognized, in which serum from some HIT patients contains subthreshold levels of platelet‐activating antibodies (by SRA) that become detectable using a PF4‐enhanced platelet activation assay. Unusual immunologic features of HIT include early antibody detectability (at onset of platelet count fall) and antibody transience (seroreversion). Widely available PF4‐dependent enzyme immunoassays (EIAs) have high sensitivity but poor specificity for HIT, although specificity is enhanced with IgG‐specific EIAs and strong positive results; unfortunately, EIA results are usually not available in real time. Automated rapid immunoassays, such as the chemiluminescence immunoassay (CLIA) and latex immunoturbidimetric assay (LIA), facilitate real‐time laboratory diagnosis. Recently available likelihood ratio (LR) data for positive (LR+) and negative (LR?) test results allow clinicians to adjust their pretest probabilities for HIT, using Bayesian analysis, into real‐time posttest probabilities that are dramatically increased (test positive) or decreased (test negative). Moreover, (semi‐)quantitative CLIA‐ and LIA‐positive results (weak, moderate, strong positive) can further refine the posttest probability of HIT.     

Analytical validation of anti-toxoplasma IgG immunoassays

There are often discrepancies when using different methods to measure anti-Toxoplasma gondii IgG levels in patient samples. The diagnostic performance of a chemiluminescent immunoassay (CLIA) and an enzyme-linked fluorescent assay (ELFA) used as confirmatory tests for samples identified as positive or equivocal by an electrochemiluminescent immunoassay (ECLIA) were examined. Cut-off values were those stated by the manufacturer, and Western blot was used to confirm the results of all methods. All samples identified as positive by ECLIA (n = 93) were confirmed as positive by Western blot, as were 14 of the 28 samples identified as equivocal. When these 121 samples were retested, the sensitivities of CLIA and ELFA were 64.4% and 73.8%, respectively. Both methods exhibited a specificity of 100%. This study confirms that the results obtained from the different immunoassays are not comparable, and neither CLIA nor ELFA should be used to confirm ECLIA results, which should instead be confirmed by methods such as Western blot or Sabin-Feldman dye test.     

化学发光法与放射免疫法分析甲状腺激素结果对比

目的通过放射免疫分析法(RIA)和微粒子化学发光免疫分析法(CLIA)对甲状腺激素的测定结果比对,探讨CLIA法在甲状腺功能测定中的临床应用价值。方法对RIA法和CLIA法分析甲状腺激素的精密度、重复性、灵敏度进行比较;采用RIA法和CLIA法,随机抽取200份血标本进行平行检测比较,结果进行相关性处理。结果测定三碘甲状腺原氨酸(T3)、甲状腺素(T4)、CLIA法批内变异系数均小于RIA法(P<0.01),测定游离三碘甲状腺原氨酸(FT3)、游离甲状腺素(FT4)和促甲状腺素(TSH),CLIA法批内变异系数低于RIA法,但无显著性差异;CLIA法测定T3、T4、FT4批间变异系数均小于RIA法(P<0.01);测定FT3、TSH,CLIA法批间变异均小于RIA法,但无显著性差异;CLIA法与RIA法均有较好的回收率;CLIA法最低检出限均低于RIA法;两法检测结果高度相关。结论CLIA法对检测T3、T4、FT3、FT4、TSH更为敏感、准确快速,精密度、重复性高于RIA法,或与RIA法接近,完全可以在临床检验中代替RIA法。     

The performance of different anti-dsDNA autoantibodies assays in Chinese systemic lupus erythematosus patients

To compare the performance of different commercial anti-dsDNA autoantibody assays, including multiplex-based immunoassay (Bio-Plex), Farr radioimmunoassay (Farr), ELISA, chemiluminescent immunoassay (CLIA), and Crithidia Luciliae indirect immunofluorescence test (CLIFT) in Chinese patients with systemic lupus erythematosus (SLE). SLE patients (n = 119) as well as healthy controls (n = 200) and disease controls (n = 100) were recruited, and serum anti-dsDNA autoantibodies were detected by Bio-Plex, Farr, two ELISA assays (Medical & Biological Laboratories-ELISA, EUROIMMU-ELISA), CLIA, and a standard CLIFT. The correlation of anti-dsDNA autoantibody levels to SLE disease activity was calculated, and the specificity and sensitivity of these methods were measured by receiver-operator characteristic (ROC) curve analysis. In ROC curve analysis, Bio-Plex showed the largest area under the curve (AUC) over other assays. Cutoff adjustment according to ROC enhanced the performance of all quantitative assays. Overall, Bio-Plex and CLIFT have higher specificity (>90.00%). ELISA and CLIA results are correlated with disease activity, and Bio-Plex results have the strongest correlation with SLEDAI score and active renal involvement. Bio-Plex assay has better overall performance in anti-dsDNA detection over Farr, ELISA, and CLIA methods in Chinese SLE patients.     

Superiority of minipool nucleic acid amplification technology for hepatitis B virus over chemiluminescence immunoassay for hepatitis B surface antigen screening

BACKGROUND AND OBJECTIVES: The Japanese Red Cross (JRC) have developed a fully automated multiplex (MPX) nucleic acid amplification technology (NAT) system for hepatitis B virus (HBV), hepatitis C virus (HCV) and human immunodeficiency virus 1 (HIV-1). This is used to test serologically negative blood units from volunteer, non-remunerated donors. The system utilizes a 50-sample pool for NAT screening with an input volume of each pool. This results in a significantly higher sensitivity for hepatitis B than that seen with highly sensitive hepatitis B surface antigen (HBsAg) testing. MATERIALS AND METHODS: From 1 February 2000 to 15 October 2001, over 11 million donations, which were serologically negative, were tested using the MPX NAT system. Donations found to be HBV DNA positive were further tested by using the chemiluminescence immunoassay (CLIA). RESULTS: Out of 181 HBV DNA-positive donations, 96 (53%) and 76 (42%) were negative by individual enzyme immunoassay (EIA) and CLIA testing, respectively. CONCLUSIONS: The sensitivity of the 50-sample pool MPX NAT system was higher than that of individual HBsAg screening by CLIA. By adopting this NAT-screening system, the JRC has improved the safety of the blood supply and maintained supply across Japan.     

Evaluation of a Particle Gel Immunoassay as a Screening Test for Syphilis

Abstract Background:   Recent trends in Western Europe show an increase in sexually transmitted infections. Surveillance data in Switzerland confirm this rising trend. Notifications of syphilis cases nearly doubled in the year 2002 and almost tripled in 2003. This trend necessitates an early correct diagnosis making reliable screening tests mandatory. Materials and Methods:   In the presented study a particle gel immunoassay (ID-PaGIA syphilis antibody test, Diamed) using recombinant treponemal antigens TpN15, TpN17 and TpN47 was evaluated as a screening test in comparison to the currently used Treponema pallidum particle agglutination test (Serodia-TPPA, Fujirebio). Serum samples were obtained from a cross-sectional sero-epidemiological study among men who have sex with men. Samples were tested with the PaGia and the TPPA. In the case of equivocal results a titrated TPPA of an external laboratory was used as a confirmation test. Results:   In total 650 serum samples (48 seropositive patients, 602 negative) were evaluated. The PaGIA showed a sensitivity of 0.89 (43/48) and the TPPA of 0.83 (40/48). This difference was not statistically relevant (p = 0.4). The particle gel assay showed a significantly higher specificity (1.0) compared to the TPPA (0.98) (p = 0.004). Conclusion:   The PaGIA showed a sensitivity comparable to that of other treponemal tests with an even better specificity. Advantages of the PaGIA are the fast reaction time of only 20 min and the simplicity of the procedure with minimal technical equipment.