Direct chemiluminescence immunoassay for estradiol in serum

In this simple, reliable, fast solid-phase chemiluminescence immunoassay for directly measuring (i.e., without prior extraction) estradiol-17 beta in serum, a monoclonal antibody is used that binds estradiol with high affinity (Ka = 10(10) L/mol), and does not bind other steroids tested, the highest cross reactivity observed being 0.1% for estradiol-17 alpha. In this system the monoclonal antibody is bound to the wells of microtiter plates via a second antibody directed against the monoclonal antibody. Fifty microliters of serum and estradiol-displacing agents are added, followed by 100 pg of estradiol-isoluminol conjugate, and the label is measured by luminometry after the binding reaction. The sensitivity of the assay is 180 pmol per liter of serum, and the effective working range at less than or equal to 10% CV is 270 to 6700 pmol/L. Analytical recovery of added estradiol averaged 99.7% (SD 6.5%). Within- and between-assay CVs ranged between 5 and 12.7%. Thirty-five unknown serum samples can be assayed within 4 h. Results correlated well with those obtained with a direct RIA: r = 0.94 (n = 149). This assay opens new perspectives for chemiluminescence immunoassays.     

Direct solid-phase enzyme immunoassay of progesterone in saliva

We have developed a rapid enzyme immunoassay for progesterone in saliva. This solid-phase assay is carried out on microtitre plates with no extraction or centrifugation steps. The detection limit of the assay is 200 fg per well (3.2 pmol/L). Intra- and interassay coefficients of variation for low, medium, and high concentrations of progesterone were 7.5, 16.0; 9.1, 8.3; and 8.7, 6.7%, respectively. Correlation between total plasma progesterone (assayed by enzyme immunoassay with extraction) and salivary progesterone concentrations was good (r = 0.848, p less than 0.001, n = 56). We found the assay useful for monitoring ovarian function. The analytical procedure is convenient, and one person can assay more than 200 saliva samples per working day. The turnaround time for 36 samples is 2 h, including 1.5 h of incubation time, when previously coated plates are used. We conclude that such assays are very suitable for measuring progesterone in serial saliva samples and could become the preferred method for monitoring ovarian function.     

Direct solid-phase chemiluminescence immunoassay for salivary progesterone

A simple, direct chemiluminescence immunoassay for progesterone in mixed, unstimulated saliva is described. We use purified polyclonal anti-progesterone antibodies covalently coupled to polyacrylamide beads and progesterone-11 alpha-hemisuccinyl-aminobutylethyl isoluminol as the chemiluminescent ligand marker. Bound and free ligand are separated by simple centrifugation. The detection limit of the assay is 1.5 pg per tube (38 pmol/L). Intra- and interassay coefficients of variation for low, medium, and high progesterone concentrations are 7.9, 6.8, 8.8% and 9.3, 6.9, 8.5%, respectively. Analytical recovery of added progesterone is 99.6%. Mean +/- SD progesterone concentrations (pmol/L) in saliva are 178 +/- 46 in the follicular phase, 313 +/- 90 in the periovulatory phase, and 658 +/- 166 in the luteal phase of the menstrual cycle. Correlation between progesterone concentrations in serum (assayed by RIA after extraction) and in saliva is good (r = 0.88, p less than 0.001, n = 96). The assay is simple, fast (4 h, including 1.5 h of incubation time), and reliable.     

电化学发光免疫法和化学发光微粒免疫分析法在HBsAg检验中的价值比较

目的比较电化学发光免疫法(CI)和化学发光微粒免疫分析法(CMIA)在乙肝表面抗原(HBsAg)检验中的价值。方法纳入我院收治的乙型肝炎疑似病例396例,分别使用CMIA、CI法进行HBsAg水平的测定。以CMIA为金标准,分析CI对HBsAg的诊断灵敏度、特异度和符合率。计算CMIA、CI检测HBsAg的检测批次内和批次间精密度。使用Bland-Altman偏差分析对CI、CMIA的诊断一致性进行分析。结果以CMIA为检测金标准,CI检测的诊断灵敏度为95.03%(287/302),诊断特异度为97.87%(92/94),诊断符合率为95.71%(379/396)。CI、CMIA对HBsAg检测的批次内变异系数和批次间变异系数比较,差异无统计学意义(P>0.05)。CI和CMIA对HBsAg诊断的组内相关系数为0.896,CI、CMIA在对HBsAg的诊断中,一致性较好。经Bland-Altman偏差分析得,两种检测方法的偏倚程度置信区间为(-6.36~23.24),CI、CMIA在0~23 IU/mL的区域内具有良好的一致性,随着机体HBsAg水平的增加,偏差逐渐上升,2.02%(8/396)的点位于偏倚程度95%置信区间以外。结论与CMIA相比,CI在对HBsAg的检验中具有良好的诊断灵敏度、符合率以及精密度。此外,CMIA、CI对HBsAg的诊断一致性良好,然而在高水平HBsAg的检测中仍然存在一定偏差。     

Homogeneous immunoassay based on chemiluminescence energy transfer

The chemiluminescent compound aminobutylethyl-isoluminol (ABEI) and its isothiocyanate derivative have been coupled to a range of haptens (progesterone, cyclic AMP, cyclic GMP) and protein antigens (IgG, C9). All the derivatives were chemiluminescent, immunologically active, and stable for more than six months. When the ABEI-labeled antigens bind to their respective fluorescein-labeled antibodies, there is a shift in the ratio of chemiluminescence at 460 nm (blue) to that at 525 nm (green). This nonradiative energy transfer was used to establish homogeneous immunoassays, which require no separation step. These assays were at least as sensitive as the conventional radioimmunoassays and could accurately measure substances in serum (i.e., for IgG, results correlated by r = 0.96 relative to those by 125I assay) and tissue extracts (cyclic AMP, r = 0.91 relative to 3H assay), and also were used to evaluate the kinetics of antibody-antigen binding. Chemiluminescence energy transfer provides a new method for quantifying ligand-ligand interactions in the 10(-15) to 10(-18) mol range without first separating bound and free ligand. This provides a unique opportunity to investigate chemical events in single cells and intact cells.     

临床化学发光免疫法检测AFP的分析性能验证与实验方法

目的建立临床化学发光免疫检验方法学性能验证方案和实验方法。方法参考美国临床和实验室标准化协会（CLSI）系列文件和相关文献，结合工作实际，设计验证方案，并对Bayer Centaur 240化学发光免疫检测系统测定血清AFP的精密度、准确度、分析灵敏度、分析测量范围、临床可报告范围和生物参考区间6大分析性能进行验证和评价，并将实验结果与厂家（德国拜尔公司）提供的分析性能或公认的质量指标进行比较。结果AFP含量在77．4、168．0ng／ml时，日间不精密度分别为5．70％和4．84％，与厂商日间不精密度（4．9％和5．0％）基本一致；4个浓度校准品检测结果与靶值的相对偏倚均〈5％，5份室间质评控制品检测结果与靶值的相对偏倚在-3．4％-11．9％之间；检测低限为1．04ng／ml，略低于厂商的检测低限（1．30ng／ml），生物检测限在2．65—3．53ng／ml之间，功能灵敏度为3．53ng／ml；分析测量范围为3．53—912．00ng／ml（厂商线性范围1．3—1000．0ng／ml），临床可报告范围为3．53—182400．00ng／ml；生物参考区间验证结果为0．6—7．7ng／ml（厂商参考区间〈8．1ng／ml）。结论Bayer Centaur 240化学发光免疫检测系统检测AFP的分析性能符合临床要求，验证方案和实验方法简便易行。本研究对规范医学实验室建设和实验室认可，提高化学发光免疫检验质量有重要意义。     

Evaluation of a fully automated procalcitonin chemiluminescence immunoassay

We evaluated a new fully automated microparticle immunoassay for procalcitonin (LIAISON BRAHMS PCT) in comparison with a previously established manual chemiluminescence assay from the same manufacturer (LUMItest PCT, BRAHMS AG). Procalcitonin (PCT) is an early and rather specific marker of systemic bacterial infection. In addition, the efficacy of antibiotic therapy can be monitored by sequential analysis of PCT values. This is why rapid and accurate determinations of PCT are urgently required by intensive care units. The aim of this study was to evaluate in a clinical set-up a new fully automated rapid PCT test. Analytical results are compared with results obtained by a previously introduced quantitative manual test. Intra-assay coefficients of variation (CV) were found in the range of 0.94 to 7.1% at concentrations between 0.46 and 97.2 microg/l. Over a time period of 27 days the inter-assay CV was found below 4.0% at concentrations of 1.93 and 14.29 microg/l and 9.9% at 0.40 microg/l. The functional sensitivity at a CV level of 20% was determined as 0.2 microg/l. Linearity could be demonstrated in a concentration range from 0 to 445 microg/l. When serum and plasma with EDTA, citrate or heparin anti-coagulation were analyzed in parallel, no systematic bias was found. A method comparison by regression analysis showed PCT values determined by both tests in very good agreement (r = 0.99). PCT concentrations in apparently healthy subjects (n =101) were below 0.58 microg/l in line with previously published results. Patients with sepsis (n = 43) or with infectious adult respiratory distress syndrome (ARDS) (n = 28) showed median values of 22.2 and 18.9 microg/l, respectively. In a clinical set-up the LIAISON Brahms PCT assay provided rapid and accurate PCT results supporting the early detection of severe sepsis, the differentiation between systemic bacterial infection and other inflammatory diseases, and the monitoring of antibiotic therapy in septic patients. The results of the new LIAISON BRAHMS PCT assay show an excellent concordance with the LUMItest PCT. The clinical information derived from the measurements is well comparable to the results obtained with the LUMItest PCT, too.     

化学发光免疫测定技术在甲状腺肿瘤中的检验价值

目的：研究分析化学发光免疫测定技术在甲状腺肿瘤中的检验价值。方法：选取浙江省人民医院毕节医院检验科在2022年8月-2024年2月期间收治的110例疑似甲状腺肿瘤患者按照随机数字表法分为观察组（n=55,采取化学发光免疫检验）和对照组（n=55,采取放射免疫分析检验）,将病理诊断结果作为金标准。对比两组的诊断结果。结果：将病理检查结果作为金标准,阳性、阴性观察组分别检出29例、22例,对照组分别检出22例、17例,观察组的敏感度、特异度、准确度均要高于对照组,差异有统计学意义（P＜0.05）;两组的实验室指标做比较,观察组的A-TPO、Tg、TSH、FT3、A-TG、FT4指标水平均要高于对照组,差异有统计学意义（P＜0.05）。结论：在甲状腺肿瘤的检验中,采用化学发光法免疫测定技术有着良好的应用优势。     

Multicenter evaluation of assays for estradiol and progesterone in saliva

Blood samples can be difficult to obtain in studies involving serial sampling, especially in developing countries where there may also be logistic, ethical, and cultural constraints that make frequent blood collection impractical. Assays for steroids in saliva may avoid some of these difficulties. A multicenter study involving laboratories in five countries was carried out to compare the results of assays for salivary estradiol and progesterone performed with centrally provided reagents and assay protocols. Concentrations of salivary steroid as obtained by all but one center were comparable with those reported in the literature. We conclude that assays of hormones in saliva are useful adjuncts to those performed on other body fluids.     

雌二醇化学发光免疫测定法的建立

目的　建立灵敏度高 ,测定范围宽的检测人血清雌二醇 (E2 )的化学发光免疫分析法 (CLIA)。方法　采用竞争抑制法 ,用碱性磷酸酶标记抗原 ,金刚烷 (CSPD)增敏化学发光体系作为酶底物。结果　敏感度为 2 .0pg/ml;在 10～ 10 0 0pg/ml之间可以准确定量 ;用不同浓度的E2 质控血清测定精密性 ,批内变异 <8% (n =2 0 ) ,批间变异 <10 % (n =2 0 )。与雌醇、雌酮、雌三醇(E3 )的交叉反应 <1% ,与睾酮、可的松等无交叉反应。试剂具有良好的稳定性 ,在 4℃保存 14个月其整体变化幅度 <10 %。与BeckmanAccessTm及其配套试剂比较有较好的相关性。结论　方法灵敏度高 ,特异性强 ,稳定性好 ,检测范围宽 ,准确性和重复性好。     

Direct solid-phase enzymoimmunoassay of testosterone in saliva

This competitive, solid-phase enzymoimmunoassay for testosterone in saliva is carried out on microtiter plates and involves no chromatographic or extraction steps. With an overnight incubation the detection limit of the assay is 230 fg per well (16.1 pmol/L). There was a good correlation (correlation coefficient 0.95) between testosterone concentrations measured with and without prior extraction of the saliva samples. Repeated assay of three control saliva samples containing a range of testosterone concentrations (200-1000 pmol/L) gave within- and between-assay coefficients of variation of 5.5-13.2%. The analytical procedure is simple and closely resembles already published procedures for the determination of progesterone and estrone (with extraction) in saliva. One person can assay 200 samples in 24 h and the assay is suitable for reproductive and sports medical applications, particularly for projects involving serial sampling and yielding large numbers of samples.     

化学发光测定TSH的实验评价及其临床应用

用"第三代"化学发光免疫分析"夹心法",评估化学发光检测TSH的实验性能.结果显示该法精密度:批内与批间CV分别为2.08～2.74%与3.85～4.43%.回收率100.6%,灵敏度0.01 mIU/L,交叉污染率＜0.01%,与RIA法对比试验Y＝0.8942X+0.2031(r＝0.9867).因此,化学发光法具高灵敏度和高特异性,使测得的TSH量不但能区分甲减,且能用于诊断甲亢,甚至亚临床型或潜在型甲状腺功能亢进或减退.     

A new modular chemiluminescence immunoassay analyser evaluated.

Thyrotropin (TSH), free thyroxine (fT4) and testosterone assays have been used as a probe to evaluate the performances of a new modular chemiluminescence (CL) immunoassay analyser, the Abbott Architect 2000. The evaluation was run in parallel on other systems that use CL as the detection reaction: DPC Immulite, Chiron Diagnostics ACS-180 and ACS Centaur (TSH functional sensitivity only). TSH functional sensitivity was 0.0012, 0.009, 0.033 and 0.039 mU/I for the Architect, Immulite, ACS Centaur and ACS-180, respectively. Testosterone functional sensitivity was 0.38, 3.7 and 2.0 nmol/l for Architect, Immulite and ACS-180, respectively. Good correlation was obtained between the ACS-180 and Architect for all assays. The Immulite correlation did not agree well with the Architect or ACS-180 for fT4 and testosterone but was in good agreement for TSH. Regarding fT4 and testosterone, equilibrium dialysis and isotopic dilution gas-chromatography mass-spectrometry (GC-MS) respectively were used as reference methods. For both within- and between-run precision, the Architect showed the best reproducibility for all three analytes (CV < 6%).     

胰岛素化学发光法测定的建立与评价

目的建立化学发光法测定胰岛素及临床糖尿病患者胰岛素治疗跟踪与稳定的可行性评估.方法CLIA采用一步免疫分析夹心法与RIA的测定结果进行比较.并将该法运用于临床非胰岛素依赖型糖尿病进行观察.结果CLIA测胰岛素的批内与批间的变异系数分别为4.8%和6.4%,平均回收率为98.6%,与RIA的相关系数为0.8505(P＞0.05).结论CLIA法测胰岛素的各技术参数优于RIA法,能及时有效地满足糖尿病患者胰岛素治疗的临床需要.     

胰岛素化学发光法测定的建立与评价

目的：建立化学发光法测定胰岛素及临床糖尿病患者胰岛素治疗跟踪与稳定的可行性评估。方法：CLISA采用一步免疫分析夹心法与RIA的测定结果进行比较,并将该法运用于临床非胰岛素依赖型糖尿病进行观察。结果：CLISA测胰岛素的批内与批间的变异系数分别为4．8％和6．4％,平均回收率为98．6％,与RIA的相关系数0．8505（P＞0．05）。结论：CLISA法测胰岛素的各技术参数优于RIA法,能及时有效地满足糖尿病患者胰岛素治疗的临床需要。