化学发光酶联免疫分析检测血清中麻疹病毒抗体

化学发光酶联免疫分析检测血清中麻疹病毒抗体章竹君，邹克渭，程明洁（陕西师范大学分析科学研究所，西安，７１００６２）（陕西省卫生防疫站）关键词麻疹病毒抗体，化学发光，酶联免疫分析，辣根过氧化物酶目前在计划免疫工作中通常采用间接酶联免疫吸附分析法（ＥＬＩ...     

化学发光酶联免疫分析测定血清中抗DNA抗体

建立起适于临床应用的抗ＤＮＡ抗体化学发光酶联免疫分析法。该法精密度良好，相对标准偏差为２．４％。比ＥＬＩＳＡ法更加简便、经济、省时，同时提高了灵敏度（８倍）和血清的阳性检出率。探讨了检测系统中对碘苯酚增强鲁米诺－过氧化氢－辣根过氧化物酶化学发光反应机理。     

电化学分析法测定硒的研究进展

本文综述了国内外电化学分析(极谱和溶出伏安法)测定痕量硒的研究进展,主要包括极谱催化波、吸附波以及阴极溶出伏安法,阳极溶出伏安法,并从反应体系、催化波类型、检出限和电极反应机理等方面进行归纳与评述,展望了硒分析方法的研究方向和发展前景,对进一步研究硒的电化学性质和探索测定硒的新体系有重要参考价值。     

单纯疱疹病毒抗原的电化学酶免疫法检测及其分型

将酶联免疫吸附试验与单扫描示波极谱法检测相联接，用自制的小型“Ｄｍｅ－Ｐｔ－Ａｇ／ＡｇＣｌ”三电极系统，与商品酶联板配套，直接检测酶促反应电活性产物，建立了ＥＬＩＳＡ－ＬＳＰ法检测单纯疱疹病毒抗原并进行分型的方法。     

增强化学发光酶免疫分析法测定血清中的铁蛋白

本文采用对碘苯酚增强的Ｌｕｍｉｎｏｌ－Ｈ２Ｏ２－ＨＲＰ化学发光反应体系作为免疫分析的最终检测手段，建立了一种新的铁蛋白的免疫分析方法，与酶联免疫分析法相比，该方法具有灵敏度高，线性范围宽等特点。方法的相对标准偏差３．７％，标准曲线线性范围为０．１２－２０００ｎｇ／ｍｌ，用该方法对血清中的铁蛋白进行了测定，其结果与ＥＬＩＳＡ法所得结果一致。     

毛细管电泳免疫分析法研究癌胚抗原与抗体相互作用

采用毛细管电泳免疫分析法研究癌胚抗原和抗体相互作用.探讨了缓冲体系、癌胚抗原和抗体的配比、进样时间,进样电压等因素对分离检测的影响.结果表明分离电压为14 kV,进样时间为10 s, 在pH值为5.92的Tris-乙酸缓冲体系（TAE）中, 癌胚抗原及其复合物得到较满意的分离.     

木糖醇示波极谱分析法的研究

本文提出,本糖醇分子经高锰酸钾在加热条件下氧化为具有电活性的物质。在pH为8.0～9.0的氨-氯化铵-亚硫酸钠体系中,用IP-Ⅱ型示波极谱仪测定,在电位为-1.7伏时,出现一个高灵敏度的极谱峰。     

吡哆醛的电化学行为研究

用单扫示波极谱法，吡哆醛在０．２ｍｏｌ／ＬＫＣｌ＋０．０２ｍｏｌ／Ｌ ｎａＯＨ底液中，产生一良好的寺阶导数峰，ＥＰ＝－１．３０±０．０１Ｖ（ＳＣＥ），其峰高与浓度在２×１０＾－７－２×１０＾－４ｍｏｌ／Ｌ范围内成线性关系，检出限为１×１０＾－７ｍｏｌ／Ｌ实验证明，其电极过程为伴有微弱吸附性质的的可逆扩散过程（ｎ＝２），不在弱酸性和中性介质中时，吡哆醛的电极过程为前行动力学过程，其电极反应机理为伴有     

穿心莲内酯的电化学研究

用单扫描示波极谱法在０．０５ｍｏｌ／ＬＮＨ４ＣＬ＋０．２５ｍｏｌ／ＬＮＨ３．Ｈ２Ｏ中，穿心链内酯有一灵敏的还原峰，２５℃时，峰电位为－１．６０Ｖ（ｖｓ．ＳＣＥ），峰电流与浓度在０．０１～１．２ｍｇ／Ｌ和１．４～２６ｍｇ／Ｌ范围内有良好的线性关系，检测限为６ｎｇ／ｍＬ。用本法测定天然中药穿心链中总内酯的含量，其结果令人满意。实验证明穿心链内酯的电极过程为可逆的逐级电子过程，Ｈ２Ｏ２可催化该还原峰，另     

Novel and Ultrasensitive Sandwich Enzyme Immunoassay (Sandwich Transfer Enzyme Immunoassay) for Antigens

Abstract

A novel and ultrasensitive sandwich enzyme immunoassay (sandwich transfer enzyme immunoassay) for antigens is described. Antigens were reacted with dinitrophenyl monoclonal mouse antibody IgG₁ and rabbit antibody Fab′-ß-D-galactosidase conjugates. The complex formed of antigens with dinitrophenyl monoclonal mouse antibody IgG₁ and rabbit antibody Fab′-ß-D-galactosidase conjugates was trapped onto affinity-purified rabbit (antidinitrophenyl bovine serum a1bumin) IgG-coated polystyrene balls. After eliminating excess of the conjugates, the complex was eluted from the polystyrene balls with dinitrophenyl-L-lysine and transfered to clean polystyrene balls coated with affinity-purified rabbit (anti-mouse IgG) IgG. ß-D-Galactosidase activity bound to the (anti-mouse IgG) IgG-coated polystyrene balls was assayed by fluorimetry. Nonspecifically bound ß-D-galactosidase activity considerably decreased with less decrease in specifically bound ß-D-galactosidase activity. As a result, the detection limits of human thyroid-stimulating hormone (0.01 nu, 0.02 amol) and human growth hormone (10 fg, 0.5 amol) by the present enzyme immunoassay were 30-fold lower than those by the conventional enzyme immunoassay, in which antigens were incubated with monoclonal mouse antibody IgG₁-coated polystyrene balls and rabbit antibody Fab′-ß-D-galactosidase conjugates.     

Novel and Sensitive Noncompetitive Enzyme Immunoassay (Hetero-Two-Site Enzyme Immunoassay) for Arginine Vasopressin

Abstract

A novel and sensitive noncompetitive enzyme immunoassay (hetero-two-site enzyme immunoassay) for arginine vasopressin in plasma is described. Plasma (0.3 ml) was diluted 1.3-fold with an appropriate buffer and filtered by centrifugation in a micro-concentrator with polysaccharide membrane to eliminate plasma proteins. Arginine vasopressin in plasma filtrates was biotinylated and trapped onto anti-arginine vasopressin IgG-coated polystyrene balls. After washing the polystyrene balls to eliminate other biotinylated substances, the biotinylated arginine vasopressin was eluted from the polystyrene balls with HCl and was reacted with anti-arginine vasopressin Fab′-peroxidase conjugate. The complex formed was trapped onto streptavidin-coated polystyrene balls. Peroxidase activity bound to the polystyrene balls was assayed by fluorometry. The detection limit of arginine vasopressin was 11 fg (10 amol)/tube. This was 45-fold lower than that by competitive enzyme immunoassay using the same antiserum as used in this study and 9 to 400-fold lower than those previously reported by competitive radioimmunoassays. The assay range of arginine vasopressin in plasma was 0.14–140 ng /l using 100 μl of plasma filtrates corresponding to 75 u1 o f plasma. Plasma levels of arginine vasopressin i n 8 healthy subjects aged 25–41 yr with, ad libitum water in take and normal activity approximately 4 h after breakfast were 0.72 ± 0.22 (SD) ng /l (range, 0.42–1.04 ng /l).     

Electrochemical Immunoassay for Free Prostate Specific Antigen (f‐PSA) Using Magnetic Beads

Prostate specific antigen (PSA) is a prominent marker for the prostate carcinoma. It is found in human blood in free (f‐PSA) and complex forms. These two forms together are called total PSA (t‐PSA). Estimation of both forms is essential to predict malignancy. In this study we report a unique and effective technique of electrochemical detection of f‐PSA using magnetic beads on a three‐electrode screen‐printed sensor. A magnetic bead enzyme linked immunosorbent assay (ELISA) was performed in a cuvette. Following the immunoassay, magnetic beads were recovered by a magnetic concentrator and transferred on the working electrode of the 3‐electrode assembly. The amperometric response, a measure of the amount of residual enzyme activity on the beads and hence the concentration of analyte in solution, was determined by addition of enzyme substrate. The device has a detection limit of <0.1 ng mL^?1 f‐PSA and a linear range of 0 to 1 ng mL^?1 f‐PSA.     

An Enzyme Immunoassay for the Determination of Methabenzthiazuron

Abstract

A sensitive enzyme immunoassay is described for the determination of the urea herbicide methabenzthiazuron. The assay is carried out with polyclonal antibodies, which were raised in rabbits by immunization with a methabenzthiazuron-BSA conjugate containing five methabenzthiazuron residues per molecule. The ELISA was optimized on microtiter plates with a peroxidase-methabenzthiazuron tracer. The middle of the test (50% B/B₀) was found at 1.0 μg/l. The lower detection limit of methabenzthiazuron is c. 0.05 μg/l. Samples can be measured up to 10 μg/l methabenzthiazuron (upper detection limit). The assay does not require concentration or clean-up steps for drinking or ground water samples. Validation experiments showed a good accuracy and precision. Work with monoclonal antibodies is in progress.     

A New Generic Enzyme Immunoassay for Sulfonamides

Abstract

The study was directed to the development of generic immunoenzyme assay for sulfonamides. N‐sulfanil‐4‐aminobutyric acid which mimics the common part of sulfonamides was used as immunogenic hapten. The obtained rabbit polyclonal antibodies allowed detection of the N‐sulfanil‐4‐aminobutyric acid in indirect competitive immunoenzyme assay down to 0.03 ng ml^?1. The lowest detection limit for the sulfonamides tested, 0.1 ng ml^?1, was observed for sulfamethoxypyridazine. Eleven other sulfonamides could be determined at concentrations ranging from 1–37 ng ml^?1. Thus, the proposed technique can be used in class‐specific sulfonamides detection in products of animal origin.     

Electrochemical Determination of Cortisol by Capillary Electrophoretic Enzyme Immunoassay

In recent years, capillary electrophoretic immunoassay (CEIA) has become a primary analytical tool in clinical diagnostics. The primary detection methods in CEIA are UV absorbance detection and laser induced fluorescence detection (LIF)1. The major disadvantages of the UV detector are the lack of sensitivity and not available for many antigens which are small molecules without strong UV absorbance. LIF is a more general approach to improve sensitivity. However, it is difficult to u…     

Liquid Chromatography with Electrochemical Detection of Phenol and NADH for Enzyme Immunoassay

Abstract

Enzyme immunoassays based on chromatographic separation and amperometric detection of an enzyme generated product have been investigated. These assays combine the selectivity of the antigen/antibody reaction with the high sensitivity of thin layer amperometry. The feasibility of utilizing LCEC as a detection scheme was demonstrated using the Syva EMIT® kit for phenytoin. NADH production by glucose-6-phosphate dehydrogenase was monitored following a homogeneous procedure. Heterogeneous assays were developed for alkaline phosphatase labeled species which were based upon LCEC determination of phenol. Assays were designed for a common serum glycoprotein (orosomucoid) and a clinically important drug (digoxin). Detection limits approach the pg/mL level and as such may prove fruitful in the quantitation of numerous antigens of clinical interest.     

增强化学发光体系酶联免疫分析法测定人绒毛膜促性腺激素

应用活化鲁米诺,用优化的增强化学发光酶联免疫分析体系测定人绒毛膜促性腺激素,检测限为0.2mIU/ml。线性范围0～200mIU/ml,与放射免分析测定结果比较,相关性良好。进而又发展了一种半定量的照相测定法,通过实际血清样品测定,效果良好。