ELISA法和化学发光法对血清中HIV-1HIV-2抗体、梅毒抗体和丙肝抗体的检测意义

目的探讨分析ELLSA法(酶联免疫吸附法)和化学发光法对血清中HIV-1/HIV-2抗体、梅毒抗体和丙抗体的临床检测意义。方法将382份脐血作为研究分析对象,分别采用酶联免疫吸附和化学发光法对所选标本进行检测,包含丙肝抗体、梅毒抗体、HIV-1/HIV-2抗体检测。结果采用ELLSA检测后HIV-1/HIV-2抗体、梅毒抗体、丙肝抗体阳性率分别为0.52%、0.79%、1.05%。采用化学发光法检测HIV-1/HIV-2抗体、梅毒抗体、丙肝抗体3种检测的阳性率分别为0.79%、1.05%、1.31%。标准抗体品梯度稀释后再进行检测,将此3种抗体稀释程度为10 pg/m L,三组抗体采用化学发光法进行检测,其结果均为阳性,采用酶联免疫吸附方式进行检测的只有梅毒抗体呈阳性,其余两项均为阴性。结论化学发光法作为临床进行血清中HIV-1/HIV-2抗体、梅毒抗体、丙肝抗体检测的首选方案,此检测方式具有更高的灵敏性。     

基于高灵敏探针海萤荧光素类似物的化学发光法检测尿微量白蛋白

海萤荧光素类似物FCLA能检测1O2并产生532nm的化学发光,白蛋白(HSA)能强烈增强FCLA-1O2化学发光。基于此,建立了一种有效、高灵敏的检测HSA的化学发光(CL)检测技术。在选定的最佳实验条件下,HSA浓度与增强化学发光信号在0.34～5.5mg/L之间呈良好的线性关系;检出限为0.017mg/L(3σ)。对比荧光光谱分析法、同步荧光扫描法及瑞利光散射法等传统方法,本方法成本低廉、灵敏度高。实际尿样检测与临床检测结果一致,同时还初步探讨了HSA增敏CL的机制。     

化学发光成像法测定人体血清中乙型肝炎病毒

基于对碘苯酚增强的luminol-H2O2-HRP化学发光反应,利用化学发光成像法检测乙肝病毒(HBV)。用该法对人体血清中的乙型肝炎表面抗原、表面抗体、e抗原、e抗体以及核心抗体进行测定,其结果与ELISA法所得结果一致,对表面抗原检测结果为阳性的病人血清测定9次,结果的相对标准偏差为4．2％。     

胶束微观非均相体系与化学发光能量转移

胶束微观非均相体系能提高化学发光的选择性和灵敏度,并且实验操作方便,因此其在化学发光检测技术中的应用已成为研究热点.胶束增敏化学发光是对激发量子效率,荧光量子效率以及反应的速率综合作用的结果.本文综述了正相和反相胶束体系在能量转移型化学发光体系中的应用,同时阐述了胶束与不同发光体系的相互作用.     

化学发光免疫分析技术在微生物检测中的应用

化学发光免疫分析技术凭借其化学发光的高灵敏性和免疫反应的高特异性在微生物快速检测中广泛应用。该文着重叙述了化学发光免疫分析技术核心检测体系在微生物检测中的研究进展,并对其涉及到的样品前处理和检测新方法进行了综述。化学发光免疫分析技术检测微生物用时短、成本低,但特异性识别方法和检测的灵敏度有待提升,新型的发光体系、发光放大方法、可替代抗体的识别分子以及相关的前处理技术是未来研究的重点。     

板式磁颗粒化学发光免疫分析测定人血清中糖类抗原125

在传统的板式化学发光免疫分析法和管式磁颗粒化学发光免疫分析法基础上,建立了人血清中糖类抗原125(CA125)的板式磁颗粒化学发光免疫分析方法.该方法以磁性微粒子作为分离固相,96孔板为反应容器,辣根过氧化物酶(HRP)催化H2O2-luminol化学发光体系作为检测体系.本法测定CA125的检测灵敏度可达2.0U/mL,线性范围为0～400U/mL.与常用的包被板化学发光免疫分析方法对比,该方法检测范围宽.与管式磁颗粒化化学发光法比较,其分析灵敏度与精密度高、线性范围、分析通量以及分析成本方面均显示了很好的优越性.采用该方法对人血清中CA125进行测定并与罗氏全自动电化学发光系统的测值结果进行了比对,两者显示了良好的相关性.     

LED光诱导化学发光法检测饮料中的核黄素

以化学发光法为基础,建立了以发光二极管(LED)诱导化学发光体系(LED-CL)检测饮料中核黄素含量的分析方法。样品溶液与鲁米诺溶液混合后由蠕动泵带出,经LED灯照射后产生化学发光,产生的化学发光信号由光电检测器检测。核黄素浓度检测线性范围为0.39~79.56μg/L(R≥0.999 7),加标回收率为99.3%~103%,可用于饮料中核黄素的检测。     

微流控芯片化学发光检测系统的设计及其应用

通过标准光刻、化学刻蚀及热键合技术制作微流控电泳芯片,在芯片上集成流通式化学发光检测池,实现样品的芯片电泳分离化学发光检测.采用双(2,4,6-三氣苯基)草酸酯(TCPO)-过氧化氢化学发光体系,通过微泵输送化学发光试剂.单酰化苯并氨基酸和单酰化肌氨酸在该系统中得以成功地分离检测,其检测限分别达到2.8和3.2 μmol/L.     

LS 6500型闪烁计数器检测化学发光的改进方法

提出了LS 6500型闪烁计数器进行化学发光检测时的改进方法,对提高化学发光检测数据的准确度,具有重要的意义。     

化学发光成像分析法定量检测人尿液中β-人绒毛膜促性腺激素

采用一种灵敏、简单的方法,可实现对β-人绒毛膜促性腺激素（β-HCG）的高通量检测。本法具有酶联免疫吸附（ELISA）法的特异性、增强化学发光法（ECL）的高灵敏度及化学发光成像分析高样品通量等优点。冷却电感耦合（CCD）成像系统被用于检测增强化学发光信号。发光强度值与β-HCG在12．5-400mIU／mL范围内呈线性关系;检出限为4mIU／mL。测定了妇女尿液中β-HCG的含量,分析结果与医院的诊断结果一致。     

On-capillary chemiluminescence detection for capillary electrophoresis with a single capillary.

On-capillary chemiluminescence detection for capillary electrophoresis with a single capillary was reported. A hole (about 30 microm diameter) was made on the capillary wall at about 50.5 cm from the inlet end. Hydrogen peroxide solution could enter the capillary from the hole, and mixed with luminol and copper(II) to produce chemiluminescence. The chemiluminescence was detected by a PMT under the hole. Several factors that influenced chemiluminescence intensity were investigated. The detection limits for luminol and N-(4-aminolbutyl)-N-ethylisoluminol (ABEI) were 1 x 10(-11) and 2 x 10(-10) mol L(-1), respectively. The method features simple construction and no dead volume.     

Clinical applications of chemiluminescence

This article reviews the clinical applications of chemiluminescence in routine testing and surveys the diverse applications of chemiluminescence in clinical research. In routine clinical testing, chemiluminescent labels (acridinium ester, acridinium sulfonamide) and detection reactions for peroxidase and alkaline phosphatase enzyme labels (luminol and adamantyl 1,2-dioxetane-based reactions, respectively) are widely used in immunoassay and nucleic acid probe assays (e.g. hybridization protection assay, Hybrid Capture^® assay). In clinical research the sensitivity, dynamic range and diversity of chemiluminescent assays has led to a vast range of applications, notably in protein and nucleic acid blotting, microarray-based assays, monitoring reactive oxygen species, and as detection reactions for substances separated by HPLC, capillary electrophoresis (CE), and flow-injection analysis.     

毛细管电泳场放大进样化学发光检测10-16mol/L水平钴(Ⅱ)

激光诱导荧光（LIF）以其灵敏度高及应用范围广等特点成为单分子检测的主要手段[1]. 但LIF除了仪器昂贵外, 还需对非荧光物质采用柱前或柱后衍生引入荧光团, 且该法易受拉曼散射、瑞利散射和溶剂噪音的影响, 产生高背景干扰. 化学发光（CL）检测灵敏度高, 光学系统简单、无须外加光源及分光系统, 背景低, 避免了杂散光和光源不稳定性的影响. 这些优异性能使其有可能成为毛细管电泳（CE）中的一种超灵敏检测手段. 关于毛细管电泳在线化学发光检测, 本研究组曾提出一种新的试剂混合模式[2], 并设计了一种新颖的检测接口[3], 降低了噪音, 显著提高了信噪比. 场放大进样在线富集技术可用于提高检测灵敏度, 其对有机化合物[4~6]及金属配合物[7]等方面的研究已有报道. 浓缩因子达到1 500[8]. 我们系统地研究了场放大进样对金属离子的富集作用后发现, 所研究的金属离子如钴（Ⅱ）、铬（Ⅲ）、铜（Ⅱ）及镍（Ⅱ）等的浓缩因子可达到104～106.     

Design and evaluation of capillary electrophoresis in dynamically coated capillaries coupled with chemiluminescence detection

A dynamic coating capillary electrophoresis coupled with a simplified on-line chemiluminescence detection system was designed and evaluated. In the proposed system, poly-vinylpyrrolidone was used as dynamic coating substance in the separation buffer to reduce the unwanted protein non-specific adsorption, which was first applied in capillary electrophoresis coupling with on-line chemiluminescence detection. In order to avoid complex processing, an ordinary plastic cuvette was modified as a three-way joint. The chemiluminescence reaction conditions and capillary electrophoresis separation conditions were investigated in detail. The results showed that the coated capillary can be injected protein samples at least 30 times continuously with good repeatability. Under optimal conditions, the chemiluminescence relative intensity was linear with the concentration of hemoglobin in the range of 4-1850 μg mL(-1) and the detection limit was 2.0 μg mL(-1) (S/N=3). The relative standard deviation of migration times and peak heights for 40 μg mL(-1) hemoglobin were 2.5% and 4.1% (n=11) respectively. Interference of matrix effects was overcome by the calibration according to standard addition methods. Afterwards, the method was validated successfully and was applied to detect the concentration of hemoglobin in the serum of haemolytic patients.     

Migration behavior of isoluminol isothiocyanate-labeled alpha-amino acids in capillary electrophoresis with an absorption/chemiluminescence dual detection system

In our previous study, we developed capillary electrophoresis with an ultraviolet absorption/chemiluminescence dual detection system. Here, to demonstrate one of the possible applications of the capable system as well as confirm the advantage, migration behavior of isoluminol isothiocyanate-labeled alpha-amino acids was examined in the capillary electrophoresis with a dual detection system. The labeled samples were first analyzed by absorption detection with an on-capillary, followed by chemiluminescence detection with an end-capillary. The system easily, rapidly, and simultaneously produced useful information concerning chemiluminescence quenching and amino group-labeling due to the presence of both absorption and chemiluminescence detections.     

Determination of clenbuterol by capillary electrophoresis immunoassay with chemiluminescence detection

A competitive immunoassay for clenbuterol (CLB) based on capillary electrophoresis with chemiluminescence (CL) detection was established. The method was based on the competitive reaction of horseradish peroxidase (HRP)-labeled CLB (CLB-HRP) and free CLB with anti-CLB antiserum. The factors affecting the electrophoresis and CL detection were systematically investigated with HRP as a model sample. Under the optimal conditions, the tracer CLB-HRP and the immunoassay complex were separated, and the linear range and the detection limit (S/N = 3) for CLB were 5.0-40 nmol l⁻¹ and 1.2 nmol l⁻¹, respectively. The proposed method has been applied satisfactorily in the analysis of urine sample.     

Rapid chemiluminescence biosensing of microcystin-LR

A rapid immunoassay for sensitive detection of microcystin-LR using a portable chemiluminescence multichannel immunosensor (CL-MADAG) was developed. The sensor device is based on a capillary ELISA technique in combination with a miniaturized fluidics system and uses chemiluminescence as the detection principle. Minimum concentrations of at least 0.2 μg L⁻¹ microcystin-LR could be unambiguously measured in a spiked buffer system as well as in spiked real water samples. A single sample analysis for detection of microcystin-LR could be accomplished in just 13 min on the CL-MADAG. Besides providing a highly reproducible, fast and easy to perform test format, one major advantage of the newly established capillary immunoassay is represented by the feasibility of an internal retrospective quality control mechanism. Finally, simultaneous CL-MADAG measurements employing our inhibition immunoassay and a sandwich ELISA could be successfully demonstrated.     

Analysis of Follicle-Stimulating Hormone by CE with Chemiluminescence Detection Based on Competitive Immunoassay

A competitive immunoassay and capillary electrophoresis with enhanced chemiluminescence detection have been used for determination of follicle-stimulating hormone (FSH) in human serum. The method is based on the competitive immunochemical reaction of FSH and horseradish peroxidase (HRP)-labeled FSH with anti-FSH, CE separation of antibody-bound and free HRP-labeled FSH, then chemiluminescence detection. The detection limit depends on the stability of the immune complex, which depends on analysis time and detector design, and on the chemiluminescence enhancer used. A unique chemiluminescence detector without dead volume or diluent effects was therefore used, and sodium tetraphenylboron was selected as the optimum enhancer. As a result sensitivity was substantially improved. Free HRP-labeled FSH and the immune complex could be separated within 15 min in alkaline borate buffer by use of a potential of 15 kV. Under the optimum conditions a calibration plot for FSH was established in the concentration range 0–100 mIU mL⁻¹; the detection limit was 0.06 mIU mL⁻¹. The concentration sensitivity achieved was 30 times better than that of ELISA.     

Systems of capillary electrophoresis in electrochemiluminescence analysis

The results obtained in different scientific centers from 2000 to July, 2009 on the analytical application of electrogenerated chemiluminiscence in systems of capillary electrophoresis are summarized; the trend of development in this field is shown. The peculiarities of the operation of tandem systems using capillary electrophoresis with detection by electrogenerated chemiluminescence are considered, including the socalled lab-on-a-chip concept; systematized data on analytes are presented. The main problems arising in the determination of substances in liquid samples and the trends in the development of instruments for capillary electrophoresis with detection by electrogenerated chemiluminescence are discussed.     

微量金属元素的毛细管电泳分析方法及应用

综述了毛细管电泳分析微量金属元素的基本原理、分离模式(CZE、MKEC、非水电泳、芯片分离等)、检测方法(紫外、荧光、化学发光、安培、电导、质谱联用技术)等的进展和该技术在环境、生物医学领域的研究与应用。引用文献94篇。