化学发光免疫分析测定EB病毒衣壳抗原IgM抗体的方法学评价

目的　对化学发光免疫分析法 (chemiluminescentimmunoassay ,CLIA)测定EB病毒衣壳抗原的IgM抗体 (EBV VCAIgM)进行方法学评价 ,同时将其对儿童传染性单核细胞增多症 (infectiousmononucleosis,IM)的诊断准确性与传统的ELISA测定相比较。方法　在全自动CLIA仪上测定 88例血清标本 ,同时用ELISA法测定。结果　CLIA测定EBV VCAIgM的灵敏度为 0 .6 4U/ml;低、中、高浓度时的批内CV分别为 4 .1%、1.7%和 1.4 % ,批间CV分别为 4 .9%、2 .8%和 2 .4 % ;回收率为 93%～ 10 7% ,理论值与实测值间的回归方程为Y =0 .0 96 7 1.0 0 93X ,相关系数r =0 .9996 (P <0 .0 0 0 1) ;ROC曲线显示CLIA测定敏感性和特异性均在 90 %以上 ,曲线下面积为 0 .992 ,显著高于ELISA(P <0 .0 5 )。结论　CLIA是目前测定EBV VCAIgM较好的方法 ,适用于临床检测 ,可提高小儿IM诊断水平     

Evaluation of a rapid and convenient method for determination of cytomegalovirus immunoglobulin M

We evaluated a method for determination of human cytomegalovirus (hCMV) immunoglobulin M (IgM) by CLIA, and analyzed its clinical value in patients with infectious mononucleosis. Serum samples from 407 participants were measured on an automatic CLIA analyzer. At the same time the serum samples were measured by enzyme-linked immunosorbent assay (ELISA). An assessment of technological quality (methodology) in diagnostic tests demonstrated that the sensitivity of CLIA was 1.0 AU/mL and the functional sensitivity was <1.6 AU/mL. The within- and between-assay imprecision values for different concentrations were all under 5%. Recoveries for both methods were 96-110%. The linear regression equation between expected values and measured values was y=0.644+0.986x, and correlation coefficient was 0.9991 (P<0.0001). The receiver operating characteristic (ROC) curve showed that both the sensitivity and specificity of CLIA surpassed 90%. The area under the curve (AUC) was 0.990, which was significantly higher than that of ELISA (P<0.05). The results indicate that CLIA is an excellent method for hCMV IgM measurement, and thus may be useful for clinical diagnoses.     

化学发光法检测血清甲胎蛋白的方法学评价

目的对化学发光免疫分析法（CUA）测定甲胎蛋白（AFP）进行方法学评价。方法用CLIA测定178例血清的AFP值，同时也用酶联免疫吸附试验（ELISA）对AFP进行测定。结果CLIA测定AFP的灵敏度为1．3ng／mL，低、中、高浓度的批内CV分别为4．2％、2．3％、1．6％；批间CV分别为4．8％、3．2％、2．5％。回收率为96％～104％，理论值与实测值的回归方程为Y=0．1427＋0．9992X，相关系数r=0．9900（P〈0．001）。ROC曲线显示CLIA测定的敏感性和特异性均在90％以上，曲线下面积为0．992，显著高于ELISA法（P〈0．01）。结论CLIA是目前测定AFP较好的方法，适用于临床对AFP进行检测，可对原发性肝癌进行明确诊断。     

浙江省HIV抗体盲样质控考核结果分析及不同品牌分析灵敏度比较

目的调查医疗机构选用不同方法学检测抗HIV抗体的能力情况,比较不同品牌和方法学对抗HIV抗体和抗原的分析灵敏度。方法定制6个不同浓度的抗HIV-1抗体标准物质,对218家实验室开展盲样质控考核,统计医院满分率、项目脱靶率和方法学脱靶率。采用3种不同稀释液对定制标准物质倍比稀释,获得低浓度抗HIV-1抗体和HIV-p24抗原标准物质,对考核中的6款化学发光品牌和3款ELISA品牌进行分析灵敏度的比较以及稀释液的影响比较。结果医院满分率为95.9%(209/218),抗HIV-1抗体项目脱靶率为1.8%,均为假阴性脱靶。ELISA、化学发光法(CLIA)和胶体金法(GICA)的脱靶率分别为0、13.3%和40.0%。在抗HIV-1抗体浓度为1 NCU/m L时3款CLIA品牌不能检出,1款CLIA品牌不能检出2 NCU/m L的抗体浓度。2款GICA品牌不能检出12 NCU/m L的抗体浓度。3、4代ELISA和3代CLIA品牌对抗HIV-1抗体检测的分析灵敏度优于部分4代CLIA品牌,而4代CLIA品牌对HIV-p24抗原的分析灵敏度显著高于4代ELISA。不同稀释液对检测结果影响存在显著差异,但不影响定性结果判断。结论 GICA检测抗HIV抗体存在较高的漏检风险。4代CLIA对HIV抗原检测分析灵敏度较高,但对抗HIV抗体检测的分析灵敏度相对弱于ELISA和3代试剂。稀释液的差异不是免疫定性分析的主要影响因素。     

化学发光免疫定量检测狂犬病毒抗体方法的建立

目的建立化学发光免疫法检测人血清中狂犬病毒抗体。方法以纯化抗原包被固相,配以碱性磷酸酶标记抗原及金刚烷胺作为发光底物,采用双抗原夹心法检测血清中抗体。结果该方法的敏感性为0.2 IU/m l,线性范围为0~200 IU/m l,变异系数<10%,回收率在90%~110%之间。结论化学发光法定量测定人血清中狂犬病毒抗体是一种灵敏、准确、重复性好,操作简单的方法,适用于临床检测。     

酶联免疫吸附试验与化学发光法检测TORCH抗体的方法学对比研究

目的 探讨酶联免疫吸附试验(ELISA)与化学发光法(CLIA)分别检测TORCH抗体的价值.方法 将2017年2月至2019年7月在该院进行孕前或孕期的致畸五项(TORCH抗体)检查的632例孕妇纳入研究,采集静脉血标本进行ELISA、CLIA检测,并采用病原体培养分离鉴定结果作为金标准,以此对比两种检查方法的阳性率...     

化学发光免疫分析法检测低水平HBsAg与其他方法的比较在质膜上简

目的 比较并分析化学发光免疫分析法、ELISA法、胶体金法三种方法检测低水平HBsAg的结果.方法 以化学发光免疫分析仪Architect i2000检测结果作为参考,将100例＜50 IU/ml标本分为3组,分别为：＜1 IU/ml,1～5 IU/ml,＞5 IU/ml.采用ELISA、胶体金法对3组标本进行检测,对测定结果进行比较分析.结果 应用化学发光免疫分析仪Ar-chitect i2000与ELISA测定低水平（＜1 IU /ml） HBsAg的差异有统计学意义,在＜1 IU /ml组,两者符合率仅为3.8％（P＜0.05）,（1～5） IU/ml组和＞5 IU /ml组符合率分别为76.5％和100.0％.Architect i2000与胶体金法在测定低水平HB-sAg的差异有统计学意义（P＜0.05）,两种方法在＜1 IU/ml组符合率为0,（1～5） IU /ml组符合率为2.9％,＞5 IU /ml组符合率为100.0％.结论 化学发光免疫分析法检测低水平HBsAg （＜5IU /ml）灵敏度明显高于ELISA和胶体金法.     

化学发光法检测梅毒螺旋体特异性抗体的实验评价

目的应用免疫印迹法（WB）比较临床常用的梅毒螺旋体明胶凝集试验（TPPA）与化学发光法（CLIA）检测血清梅毒螺旋体特异性抗体的符合率。方法收集7 805份临床检测的血清标本,分别用TPPA和CLIA进行梅毒螺旋体特异性抗体检测,对结果不一致的标本应用WB检测确证。结果 7 805份血清标本中,CLIA检测阳性310例,TPPA检测阳性262例。对结果不一致的48例标本应用WB检测验证,结果证实阳性36例,临界阳性8例,阴性4例,而TPPA的结果均为阴性。TPPA检测总符合率为99.44%,CLIA检测总符合率为99.95%。结论 CLIA敏感性优于临床常用的TPPA,且具有结果客观、易分析、重复性好等优点,但也存在个别假阳性和钩状效应,因此应结合TPPA及临床资料确诊。     

Enhancement of the sensitivity of a chemiluminescent immunoassay for estradiol based on hapten heterology

Objective: To develop a highly sensitive chemiluminescent immunoassay (CLIA) that measures the serum estradiol (E2) levels in postmenopausal women.

Methods: The previously developed competitive CLIA for E2 consisted of an immobilized antigen and a labeled antibody with an N-functionalized acridinium ester that was modified to enhance its sensitivity. The modifications were changing the immobilized antigen from E2 to its analogues with its expected effect on hapten heterology and selecting optimal incubation conditions.

Results: The hapten heterology in which estriol was used as the immobilized antigen instead of E2 enhanced the sensitivity of the CLIA about 3-fold. A low incubation temperature and a long incubation time also effectively increased the sensitivity of the CLIA. The combination of these modifications enhanced sensitivity about 10-fold. The proposed CLIA with a 16 pmol/L detection limit, was about 5-fold more sensitive than commercially available immunoassay kits for E2.

Conclusion: The proposed CLIA is sensitive enough to measure serum E2 levels in postmenopausal women.     

人血清胱抑素C化学发光免疫分析检测系统的建立

目的 建立可用于人血清胱抑素C(Cys C)检测的化学发光免疫分析(CLIA)系统.方法 制备抗人Cys C多克隆及单克隆抗体,以单克隆抗体包被发光板,以辣根过氧标记多克隆抗体作为游离抗体,以夹心法为检测原理,建立Cys C CLIA检测系统;对自建检测系统进行方法 学评价,并与罗氏MODULAR P800颗粒增强透射免疫分析(PETIA)系统进行比对.结果 自建CLIA检测系统线性范围为0.05~20 mg/L,灵敏度为0.05 mg/L,标准曲线线性相关系数为0.999 7,批内、批间变异系数均小于5%;与罗氏MODULAR P800 PETIA检测系统检测结果 的相关系数为0.980 2.结论 成功建立人血清Cys C CLIA检测系统,该检测系统灵敏度高、精密度好、稳定性高,能够满足临床应用要求.     

Comparison of three automatic chemiluminescent immunoassays for monitoring dynamic profile of SARS‐CoV‐2 IgG and IgM

BackgroundSeldom performance evaluation and diagnosis comparison studies were reported for different chemiluminescent immunoassay (CLIA) kits approved under an emergency approval program for SARS‐CoV‐2 infection.MethodsA total of 100 and 105 serum separately from non‐infected populations and COVID‐19 patients were detected with SARS‐CoV‐2 IgM and IgG kits. The characteristics including precision, functional sensitivity, linearity, and accuracy were evaluated for Axceed, iFlash, and Maglumi CLIA kits.ResultsMaglumi and iFlash had the best analytical sensitivity for IgM and IgG, respectively. Axceed kits had a linearity response in the detected concentration. The clinical sensitivity of Axceed, iFlash, and Maglumi was 68.0%, 64.9%, and 63.9% with a specificity of 99.0%, 96.0%, and 100% for IgM, 85.6%, 97.9%, and 94.8% with a specificity of 97.0% for IgG. ROC analysis indicated all kits had a diagnostic power greater than 0.9. Notably, either IgM or IgG kits obtained a poor agreement (Kappa value from 0.397 to 0.713) with others. Among 38 recovered patients, 94.7% had an effective immune response, and both seropositive IgM and IgG accounted for the biggest proportion (medium, 42 days onset), then followed by the single seropositive IgG (medium, 50 days onset) in Ab profile.ConclusionThe performance of CLIA kits satisfied the diagnosis of SARS‐CoV‐2 infection. Both positive of IgG and IgM contributes to improve the specificity, and a positive one will enhance the sensitivity.     

Evaluation of the Virclia® automated chemiluminescent immunoassay system for diagnosing pneumonia caused by Mycoplasma pneumoniae

Background

Mycoplasma pneumoniae is considered an important etiologic agent of community‐acquired pneumonia (CAP) in outpatients. We aimed to evaluate the diagnostic accuracy of a quick automated chemiluminescent immunoassay (CLIA) for M. pneumoniae in a population‐based prospective study of CAP.

Methods

A total of 137 outpatients diagnosed with CAP were included in the study. Acute‐ and convalescent phase sera were analyzed for IgG and IgM to M. pneumoniae with both CLIA (VirClia^®) and ELISA immunoassays. Conventional serological criteria by quantitative ELISA were considered as reference standard. Sensitivity and specificity of the assay were assessed with the construction of receiver operating characteristic (ROC) curves, and the kappa index was used to evaluate the accuracy of the IgG and IgM determinations in the acute phase.

Results

Thirty‐eight patients were diagnosed with pneumonia by M. pneumoniae. ROC curves for IgG and IgM of convalescent and acute phase (C/A) quotients by the CLIA and ELISA assays were comparable. Specifically, for the CLIA, the best C/A quotient for IgG was 2.617 (sensitivity, 94.9%; specificity, 99.9%), and for IgM 1.400 (sensitivity, 65.8%; specificity, 100%). Regarding the acute phase, the best diagnostic accuracy for the CLIA was obtained with an IgG index of 1.120 (sensitivity, 89.5%; specificity, 73.7%). The CLIA was very simple to execute and required a minimum sample handling.

Conclusion

The accuracy of the Virclia^® assay for the diagnosis of M. pneumoniae infection in outpatients with CAP was equivalent to the quantitative ELISA. The CLIA was quicker to perform and displayed better analytic workability than conventional ELISA.

     

The rapid diagnosis of infectious mononucleosis using an ELISA that detects IgM antibody to a peptide component of Epstein-Barr virus nuclear antigen.

An enzyme-linked immunosorbent assay (ELISA) that detects IgM antibody to a peptide component of the Epstein-Barr virus (EBV) nuclear antigen (EBNA-1) was compared with a conventional rapid heterophil antibody method for the rapid diagnosis of infectious mononucleosis. Discrepancies between the two methods were further analyzed using an indirect immunofluorescence assay to detect antibodies to EBV antigens. We evaluated 298 cases of suspected infectious mononucleosis. The ELISA was very sensitive (98.7%) and able to detect some cases (seven (9%) of 75 confirmed positives) that were negative by the rapid heterophil antibody test, but confirmed by immunofluorescence. However, approximately 17% of all positive tests could not be confirmed by EBV-specific immunofluorescence; thus, the overall positive predictive value was 83%; negative predictive value was 99.5%; and specificity was 93%. The high rate of false-positive tests makes this rapid ELISA unsuitable for the diagnosis of infectious mononucleosis.     

化学发光法与ELISA法检测梅毒抗体的一致性比较

目的研究化学发光法与酶联免疫吸附试验(ELISA)法检测梅毒螺旋体特异性抗体的一致性比较。方法分别用ELISA、梅毒螺旋体明胶颗粒凝集试验(TPPA)、微粒子化学发光法(CMIA)检测120例送检梅毒的血清样本。结果 ELISA和TPPA检测梅毒血清结果一致性较好(κ=0.966,P<0.05);CMIA和TPPA检测梅毒血清结果一致性好(κ=0.867,P<0.05);ELISA与TPPA一致性较化学发光法稍差。结论 ELISA和CMIA均与TPPA具有较好的相关性,CMIA敏感性最高,梅毒血清学试验检测可采用CMIA为筛查试验,TPPA作为确证试验,以减小梅毒的漏诊率和假阳性率。     

化学发光酶免疫分析法及ELISA检测抗-HCV的结果比较

目的对比化学发光酶免疫分析法及酶联免疫吸附试验(ELISA)用于丙型肝炎抗体检测的效果。方法选取丙型肝炎患者140例,抽取血液标本后分别进行丙型肝炎病毒(HCV)相关抗体的化学发光酶免疫分析法及ELISA法检测。结果化学发光酶免疫分析法检出抗-HCV阳性的检出率为98.6%,高于ELISA法的检出率(94.3%),差异有统计学意义(P0.05)。化学发光酶免疫分析法对于抗AMA-M2抗体、抗3E抗体、抗SP100抗体、抗PML抗体、抗GP210抗体的检测阳性率分别为80.0%、73.6%、37.9%、55.7%和47.9%,而ELISA法检测结果分别为12.9%、12.9%、13.6%、10.7%和6.4%,差异均有统计学意义(P0.05)。结论相对于ELISA法,化学发光酶免疫分析法在丙型肝炎中的应用有较高的检出阳性率,对于相关抗体检测有较高的敏感性,值得推广应用。     

Diagnostic value of antibodies to modified citrullinized vimentin in early rheumatoid arthritis

A hundred and two patients (18 males, 84 females; mean age, 50.3 +/- 12.3 years) diagnosed as having early rheumatoid arthritis (RA) were examined. A control group consisted of 189 patients with various rheumatic diseases and 30 healthy donors. The serum concentrations of antibodies to modified citrullinized vimentin (MCVA) and to cyclic citrullinized peptide2 (CCTP2A) were measured by enzyme immunoassay (EIA); rheumatoid factor (RF) IgM was determined by nephelometric immunoassay. In early RA, the level of MCVA (median, 49.6 U/ml; interquartile range, 0.9-249.3) was significantly higher than in the control group 1.65 U/ml; 0.3-19.7). There was a direct significant correlation between the levels of MCVA and CCTP2A (p = 0.9), as well as RF IgM (p = 0.6). The diagnostic efficiency of MCVA (area under the curve, 0.705; 95% confidence interval, 0.607-0.803) was higher than that of CCTP, (0.590; 0.467-0.714), but lower than that of RF IgM (0.813; 0.736-0.889). MCVA was comparable with CCTP, and RF IgM in sensitivity; however, it ranked below them in specificity (71%). Choice of the optimum upper normal range (30 U/ml) permits up to an 88% increase in MCVA specificity and the concurrent consideration of results of testing MCVA, CCTP2A, and RF IgM is attended by up to a 78% increase in sensitivity. EIA of MCVA is a sensitive and specific serological test for the diagnosis of early RA.     

Epidemiological study using IgM and IgG antibody titers against SARS-CoV-2 in The University of Tokyo,Japan (UT-CATS)

IntroductionThe worldwide pandemic of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has continued to date. Given that some of the patients with coronavirus disease 2019 (COVID-19) are asymptomatic, antibody tests are useful to determine whether there is a previous infection with SARS-CoV-2. In this study, we measured IgM and IgG antibody titers against SARS-CoV-2 in the serum of asymptomatic healthy subjects in The University of Tokyo, Japan.MethodsFrom June 2020, we recruited participants, who were students, staff, and faculty members of The University of Tokyo in the project named The University of Tokyo COVID-19 Antibody Titer Survey (UT-CATS). Following blood sample collection, participants were required to answer an online questionnaire about their social and health information. We measured IgG and IgM titers against SARS-CoV-2 using iFlash-SARS-CoV-2 IgM and IgG detection kit which applies a chemiluminescent immunoassay (CLIA) for the qualitative detection.ResultsThere were 6609 volunteers in this study. After setting the cutoff value at 10 AU/mL, 32 (0.48%) were positive for IgG and 16 (0.24%) for IgM. Of six participants with a history of COVID-19, five were positive for IgG, whereas all were negative for IgM. The median titer of IgG was 0.40 AU/mL and 0.39 AU/mL for IgM. Both IgG and IgM titers were affected by gender, age, smoking status, and comorbidities.ConclusionsPositive rates of IgG and IgM titers were relatively low in our university. Serum levels of these antibodies were affected by several factors, which might affect the clinical course of COVID-19.     

Sm~(3+)标记抗心磷脂抗体IgM时间分辨荧光免疫分析法的建立

目的建立高灵敏度、宽量程的定量检测患者血清中的抗心磷脂抗体(anticardiolipin antibody,ACA)IgG的Sm3+标记时间分辨荧光免疫分析方法。方法采用ACA抗原(心磷脂+β2糖蛋白I)和兔抗人IgM抗体分别作为固相抗原和钐标记抗体,如果样本中存在ACA抗体,则形成ACA抗原-ACA抗体-钐标记兔抗人IgM抗体复合物,加入解离增强液解离铕离子,检测荧光强度,样本中ACA-IgM抗体含量与荧光强度成正比;对建立的时间分辨荧光免疫(time-resolved fluoroimmunoassay,TRFIA)法检测ACA抗体的线性范围、精密度、检测范围进行分析;对25例ACA-IgM抗体阳性血清标本分别利用TRFIA及ELISA法检测ACA-IgM抗体,分析其相关性;收集50例健康献血者,利用TRFIA检测其血清中的ACA-Ig抗体,计算临床特异度。结果利用SPSS 13.0统计软件进行统计学分析,TRFIA法检测高、中、低3种浓度混合血清的批内(n=20)精密度分别为2.39%、3.26%和3.94%,批间(n=8)精密度分别为2.92%、3.73%和4.35%;方法的灵敏度为0.1 MPL U/ml;TRFIA法检测健康献血者,临床特异度为98%;TRFIA与ELISA 2种方法检测结果的一致性检验采用相关性分析,相关系数为0.956;将1份ACA-IgM抗体强阳性的血清标本从2483 MPL U/L倍比稀释至0.151 MPL U/L,ELISA方法较好的线性范围在4.85-310.4MPL U/L,而TRFIA方法在0.151-2483MPL U/L都有较好的线性。结论首次建立稳定的高灵敏度和宽检测范围的TRFIA法检测人血清中的ACA-IgM抗体,对早期诊断自身免疫性疾病及监测疗效具有重要意义,该方法以其优势有望在各检验科室得以普遍应用。     

不同方法学检测乙型肝炎血清标志物结果的评价分析

目的对酶联免疫吸附试验（ELISA）、时间分辨免疫荧光法（TRFIA）、化学发光法（CLIA）、电化学发光法（ECLIA）检测乙型肝炎血清标志物[乙型肝炎表面抗原（HBsAg）、乙型肝炎表面抗体（抗HBs）、乙型肝炎e抗原（HBeAg）、乙型肝炎e抗体（抗HBe）、乙型肝炎核心抗体（抗HBc）]的结果进行分析,评价不同方法学检测结果之间是否具有一致性。方法用ELISA、TRFIA、CLIA、ECLIA分别检测100例乙型肝炎病毒（HBV）感染者和50名正常体检者的乙型肝炎血清标志物。以ECLIA作为参考方法,通过Kappa检验评价不同方法学检测结果之间的一致程度。结果 4种方法检测乙型肝炎血清标志物的结果具有高度以上一致性,CLIA和ECLIA在检测敏感性上更有优势。结论目前广泛使用的检测乙型肝炎血清标志物的方法具有较好的一致性,为实验室结果互认提供了依据。其中ELISA适用于人群初筛,CLIA和ECLIA对于肝炎患者疗效判断更有价值。