化学发光酶免疫法测定血清地高辛

目的评价化学发光酶免疫法测定地高辛血药浓度.了解临床地高辛的应用状况.方法采用该法测定地高辛标准品及临床慢性心功能不全、正接受地高辛治疗者血药浓度与14个生化指标比较分析.结果该法批内、批间精密度分别为2.58%和5.13%,浓度处在治疗范围者仅26/71,偏低组36/71,偏高组9/71.BUN、CK、AKP在血药浓度低、中、高三组间P＜0.01,LDH、Cr、K+、Na+组间P＜0.05.BUN、AKP、K+正常组与增高组间血药浓度P＜0.05,SDCs偏高组中AKP、LDH、CK含量与对照组比较均P＜0.01,且SDCs与AKP的相关系数r=0.6737.结论化学发光酶免疫法是一种快速、可靠、准确、有效的血药监测法,可作为判断药物疗效的客观指标,肾功能减退及电解质失衡可影响血药浓度.地高辛使用可影响到骨骼肌,其中以AKP升高较为明显.     

化学发光酶免疫法测定血清地高辛

目的 评价化学发光酶免疫法测定地高辛血药浓度。了解临床地高辛的应用状况。方法 采用该法测定地高辛标准品及临床慢性心功能不全、正接受地高辛治疗者血药浓度与14个生化指标比较分析。结果 该法批内、批间精密度分别为2．58％和5．13％,浓度处在治疗范围者仅26／71,偏低组36／71,偏高组9／71。BUN、CK、AKP在血药浓度低、中、高三组间P＜0．01,LDH、Cr、K^ 、Na^ 组间P＜0．05。BUN、AKP、K^ 正常组与增高组间血药浓度P＜0．05,SDCs偏高组中AKP、LDH、CK含量与对照组比较均P＜0．01,且SDCs与AKP的相关系数r=0．6737。结论 化学发光酶免疫法是一种快速、可靠、准确、有效的血药监测法,可作为判断药物疗效的客观指标,肾功能减退及电解质失衡可影响血药浓度。地高辛使用可影响到骨骼肌,其中以ADP升高较为明显。     

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。     

ACCESS全自动微粒子化学发光免疫分析仪的使用探讨

目的：了争ＡＣＣＥＳＳ全自动微粒子化学发光免疫分析仪的主要特点及性能，对该仪器作了评价。方法：使用新鲜混合血清及质控品对分别用“三明治夹心”的促甲状腺素（ＴＳＨ）、人绒毛膜促性腺激素（β－ＨＣＧ）、及“竞争抑制”的四碘甲状腺原氨酸（ＴＴ４）、三碘甲状腺原氨酸（ＴＴ３）酶免疫微粒子化学发光法检测项目，以双盲法作评价。结果：三种不同浓度新鲜混合血清的β－ＨＣＧ、ＴＳＨ、ＴＴ３、ＴＴ４批内精密度均小于５     

Immunochromatography and chemiluminescent enzyme immunoassay for COVID-19 diagnosis

IntroductionThe rapid and accurate detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is required to prevent the spread of COVID-19. This study evaluated the utility of two SARS-CoV-2 antigen detection methods.MethodsWe evaluated two types of antigen detection methods using immunochromatography (Espline) and quantitative chemiluminescent enzyme immunoassay (Lumipulse). RT-PCR was performed as a standard procedure for COVID-19 diagnosis. Lumipulse and RT-PCR were performed for all 486 nasopharyngeal swabs and 136 saliva samples, and the Espline test was performed for 271 nasopharyngeal swabs and 93 saliva samples.ResultsThe sensitivity and specificity of the Espline test were 10/11 and 260/260 (100%), respectively for the nasopharyngeal swabs and 3/9 and 84/84 (100%), respectively for the saliva samples. High sensitivities for both saliva (8/9) and nasopharyngeal swabs (22/24) were observed in the Lumipulse test. The specificities of the Lumipulse test for nasopharyngeal swabs and saliva samples were 460/462 (99.6%) and 123/127 (96.9%), respectively.ConclusionThe Espline test is not effective for saliva samples but is useful for simple and rapid COVID-19 tests using nasopharyngeal swabs because it does not require special devices. The Lumipulse test is a powerful high-throughput tool for COVID-19 diagnosis because it has high detection performance for nasopharyngeal swabs and saliva samples.     

Multi-site analytical evaluation of a chemiluminescent magnetic microparticle immunoassay (CMIA) for sirolimus on the Abbott ARCHITECT analyzer

Objective

This study evaluated a new chemiluminescent magnetic microparticle immunoassay (CMIA) for sirolimus on the ARCHITECT analyzer.

Design and methods

Patient and laboratory proficiency samples were tested at three European sites and one site in the United States.

Results

The CMIA total %CV's were all < 8% and the Limit of Quantification (LOQ) was < 1.52 ng/mL across the four sites. It cross-reacts to sirolimus metabolites F4 and F5 and showed no hematocrit interference over a range of 25% to 55%. Patient specimen correlations to three LC/MS/MS methods gave R ≥ 0.91 at three sites and mean biases of 14%, 25% and 39%. CMIA patient specimen correlations to the Abbott IMx gave R ≥ 0.94 at 2 sites and mean biases of 5.4% and 6.9%.

Conclusions

CMIA is a precise and sensitive immunoassay method without hematocrit interference. It correlates well to both LC/MS/MS and immunoassay results, but shows an expected positive bias to LC/MS/MS.     

Rapid and sensitive chemiluminescent enzyme immunoassay for measuring tumor markers.

We developed a chemiluminescent enzyme immunoassay (CLEIA) to quantify such tumor markers as carcinoembryonic antigen (CEA), alpha-fetoprotein (AFP), CA19-9, and CA125. We used a novel chemiluminescent substrate, a derivative of 1,2-dioxetane phosphate (AMPPD), to measure alkaline phosphatase as a labeling enzyme to Fab' fragments of antibody. Regardless of the solid phase, i.e., polystyrene beads (6 mm diameter) or ferrite-coated particles (0.3 microns diameter), the standard curves within the dynamic ranges of the conventional RIA or enzyme immunoassay (EIA) were linear in all cases except for those with AFP. Use of the ferrite particles further shortens the immunoreaction, so the assay can be performed in 30 min. In addition, the relationships between concentrations of the marker and chemiluminescent signals for CA19-9, CA125, and CEA were linear up to concentrations about 10-fold greater than the ordinary dynamic ranges. Intra- and interassay CVs (averages for individual analyte) were 2.2%-4.9% and 2.0%-5.8%, respectively. In an analysis of serum samples, results of the CLEIA correlated reasonably well with those of RIA or EIA. The lower limit of detection by CLEIA with ferrite particles was 390 arb. units/L for CA19-9, 990 arb. units/L for CA125, 0.06 micrograms/L for CEA, and 0.03 micrograms/L for AFP. Thus, the sensitivity increased to between two- and 10-fold that of RIA or colorimetric EIA, depending on the analytes.     

Calcitonin levels in normal individuals with new highly sensitive chemiluminescent enzyme immunoassay

Human serum calcitonin concentration in normal individuals was measured with a new assay based on the chemiluminescent enzyme immunoassay (CLEIA) method. The CLEIA assay was highly sensitive and was able to determine a calcitonin concentration of 0.04 pg/ml as sensitivity limit at a condition of 0+3SD. With this CLEIA assay, the mean value of calcitonin in males and females was 2.26 and 1.33 pg/ml, respectively, highlighting a significant difference between genders. The mean value and range of human serum calcitonin in this assay were approximately 1/10 those reported previously in competitive radioimmunoassay (RIA) methods. Since RIAs for calcitonin showed much variability at a low concentration range due to the competitive format, they seemed to lack the necessary sensitivity to cover the normal range and appeared only useful for hyper-calcitonin phenomenon in diseases such as medullary thyroid carcinoma. The CLEIA for calcitonin provided a lower detection limit than normal range, and it can therefore be assumed that it could be applied for the measurement of hypo-calcitonin phenomena typically found in some disorders such as osteoporosis. J. Clin. Lab. Anal. 12:218–222, 1998. © 1998 Wiley-Liss, Inc.     

促黄体生成素定量测定试剂盒(化学发光免疫分析法)的临床性能评价

目的 对广州市达瑞生物技术股份有限公司的促黄体生成素定量测定试剂盒(化学发光免疫分析法)进行临床应用研究及临床性能评价. 方法 采用平行、盲法、对照试验设计,用达瑞生物公司试剂盒检测335例样本,以雅培公司的促黄体生成素测定试剂盒(化学发光微粒子免疫检测法)作为对照试剂盒,罗氏公司的促黄体生成素检测试剂盒(电化学发光法)作为复核试剂盒.操作按各试剂盒说明书进行,对达瑞生物公司试剂盒的符合率和相关性进行计算分析. 结果 同雅培公司试剂盒比较,达瑞生物公司试剂盒检测血清样品异常符合率为95.8％,正常符合率为95.2％,总符合率为95.4％,Kappa值为0.898 (P＜0.01).线性回归方程为Y=0.358+0.961X,两者测定值接近,相关系数为0.988(P＜0.01),相关性良好.同血清检测结果相比,血浆检测结果Kappa值为1.000(P＜0.01),线性回归方程为Y=0.120+0.966X,相关系数r值为0.999 (P＜0.01),相关性良好.特异性样本检测结果均为正常. 结论 本试剂盒检测性能可以满足临床应用的需要.     

人心肌肌钙蛋白Ⅰ化学发光免疫分析方法的建立

目的　建立检测人心肌肌钙蛋白I(cTnI)化学发光免疫分析方法。方法　用重组cTnI免疫新西兰兔获得抗体 ,经DEAE 2 3层析和 (NH4) 2 SO4沉淀纯化后 ,用碳化二亚胺 (EDC)法标记辣根过氧化物酶 ,SephadexG 2 0 0纯化。优化鲁米诺 H2 O2 肉桂酸发光体系 ,建立了cTnI化学发光免疫分析法。用该法检测 138例正常人及病人血清标本 ,确定正常值参考范围 ,并与ELISA法比较。结果　所得抗体的滴度为 1∶80 0 0。优化后的发光体系为 :鲁米诺 (4 .5mmol/L) ,H2 O2 (7.5mmol/L) ,四苯硼钠 (0 .8mmol/L) ,肉桂酸 (0 .4mmol/L) ,比单独用肉桂酸或四苯硼钠作增强剂灵敏度分别增加了 1.19和 1.0 5倍。本法检测灵敏度为0 .2ng/ml,线性范围为 0 .4～ 5 0ng/ml,批内CV平均 9.0 % ,批间CV平均 11.8% ,回收率为 10 4 .2 %。Hb浓度在 0～ 5 0nmol/L ,T Bil在 0～ 15 μmol/L ,TG在 0～ 2 .0mmol/L对测定结果无影响。与ELISA比较 ,两法结果一致 (P >0 0 5 ) ,相关系数r=0 .985 2(P <0 .0 1)。用BioCheck标准品为校正物 ,确定临床诊断参考范围为 <2 .12ng/ml,诊断符合率为 96 .2 %。结论　建立的cTnI化学发光免疫分析法可常规用于临床检验。     

Novel homogeneous enzyme immunoassay: Chitinase activity enhancement immunoassay (caeia)

A novel homogeneous enzyme immunoassay has been developed that takes advantage of the observation that the activity of a bacterial chitinase could be enhanced by the binding of a ligand conjugate of a monoclonal antibody to the enzyme (L-MAbchi). The enhancement was suppressed by an antiligand antibody (AbL). Free ligand molecules competed for AbL combining with L-MAbchi, and the suppression of the enhancement was reduced. Thus the concentration of free ligand could be estimated. Biotin was used as a ligand in this model experiment. When this monoclonal antibody was conjugated with a ligand (e.g., biotin), we furthermore found that the enhancement could be suppressed by an antiligand antibody (AbL). When free ligand molecules are added to the system, competition with L-MAbchi results to reduce the suppression of enhancement.©1995 wiley-Liss, inc.     

Novel and sensitive enzyme immunoassay (immune-complex-transfer enzyme immunoassay) for alpha-fetoprotein from hepatocellular carcinoma.

Alpha-fetoprotein in sera from patients with hepatocellular carcinoma was fractionated into three peaks by affinity chromatography on a column of Lens culinaris agglutinin-Sepharose 4B. One peak (the first peak), which passed through the column without adsorption, was found in both healthy subjects and patients with liver cirrhosis and hepatocellular carcinoma. The second and third peaks were reactive with L. culinaris agglutinin and found only in patients with hepatocellular carcinoma. For alpha-fetoprotein in the second and third peaks, a novel and sensitive enzyme immunoassay (immune-complex-transfer enzyme immunoassay) was developed. Alpha-fetoprotein in test serum was reacted with dinitrophenyl affinity-purified anti-alpha-fetoprotein IgG, and the complex formed was trapped onto affinity-purified (antidinitrophenyl bovine serum albumin) IgG-coated polystyrene balls. The polystyrene balls were washed to eliminate substance(s) other than alpha-fetoprotein in the test serum, and the complex was eluted from the polystyrene balls with dinitrophenyl-L-lysine. The eluted complex containing alpha-fetoprotein in the second and third peaks was trapped onto L. culinaris agglutinin-coated polystyrene balls and reacted with affinity-purified anti-alpha-fetoprotein Fab'-beta-D-galactosidase conjugate. Beta-D-galactosidase activity bound to the polystyrene balls was assayed by fluorimetry. The maximal volume of serum that could be used without interference was 20 microliters, which was 100-fold larger than that in the previous enzyme immunoassay.(ABSTRACT TRUNCATED AT 250 WORDS)     

Suitability of chemiluminescent enzyme immunoassay for the measurement of blood tacrolimus concentrations in rheumatoid arthritis

ObjectivesThe aim of this study was to evaluate the suitability of chemiluminescent enzyme immunoassay (CLIA) for the monitoring of whole-blood tacrolimus concentrations in rheumatoid arthritis (RA) patients.Design and methodsSixty-three RA patients and 47 renal transplant (RT) patients treated with tacrolimus were enrolled. Tacrolimus concentrations in spiked blood and patient blood were measured by CLIA and HPLC-MS/MS. The cross-reactivity in CLIA was evaluated using 13-O-demethylated or 31-O-demethylated tacrolimus.ResultsTacrolimus concentrations measured by CLIA correlated with those measured by HPLC-MS/MS. Bland–Altman analysis revealed the 95% confidence intervals between CLIA and HPLC-MS/MS in RA and RT patients were ? 20.7 to 109.9% and ? 5.0 to 74.1%, respectively. While 31-O-demethylated tacrolimus cross-reaction amounted to an equivalent of 120% tacrolimus in CLIA, 13-O-demethylated tacrolimus did not cross-react.ConclusionCLIA values should be carefully interpreted in RA patients, especially those receiving a low dose of tacrolimus.     

Development of a rapid and sensitive magnetic chemiluminescent enzyme immunoassay for detection of luteinizing hormone in human serum

ObjectivesA rapid and sensitive magnetic particles (MPs)-based chemiluminescence enzyme immunoassay (MPs-CLEIA) for determination of luteinizing hormone (LH) in human serum was developed and validated.Design and methodsThe method was developed based on a sandwiched immunoreaction format, in which MPs served as both the solid phase and separator. The horseradish peroxidase(HRP)–luminol–H₂O₂ chemiluminescent system was chosen as the detection system.ResultsThis method had a detection limit of 0.2 mIU/mL and a linear range of 0.5–200 mIU/mL. The assay showed little cross-reactivity, and the intra- and inter-assay coefficients of variation were less than 10%. The recoveries were from 101.6% to 104.1%. The method has been successfully applied to detection of LH in human serum and showed good correlation compared with radioimmunoassay (RIA).ConclusionThe MPs-CLEIA provided apparent advantages over RIA, and facilitated the development of high-throughput screening and automated operation systems in clinical diagnosis.     

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。     

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。     

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。