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IgG4酶联免疫分析的建立及初步应用

Development and Primary Application of IgG4 Enzyme Linked Immunoassay
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摘要 用兔抗人IgG4的多克隆抗体(IgG4Ab)包被成固相板,以识别结合待测标本中的IgG4。用生物素标记IgG4Ab得BioIgG4Ab;用亲和素标记辣根酶得HRPA。在已包被IgG4Ab的固相板中加入IgG4标准品(或待测样品),反应、洗涤后,加入BioIgG4Ab,反应、洗涤后,加入HRPA,反应、洗涤后,板上形成IgG4Ab—IgG4—BioIgG4Ab—HRPA复合物,加酶底物显色,用酶标仪在490nm波长测定OD值,作标准曲线,根据标准曲线,查出标本中IgG4含量。该法测定范围:2.5~200ng/mL,最低检出量2.1ng/mL,批内和批间CV分别8.4%和11.2%。测得血清IgG4含量:青年人(30名)为37.7±2.3ng/mL;30名献血员为42.7±9.9ng/mL,49例重症监护患者明显升高为71.2±9.2ng/mL;49例肺部感染患者明显降低为28.4±9.2ng/mL。结果表明该法稳定,灵敏度适于检出人血清IgG4水平。 Rabbit-anti-human IgG4 polyclonal antibody (IgG4Ab) was coated on solid plate in order to recognize and bind IgG4 in the samples. Another IgG4Ab was labeled with biotin to form Bio-IgG4Ab, and horse radish peroxidase was labeled with avidin to form HRP-A. Adding IgG4 standards or samples on the coated plate after reaction and washing, Bio-IgGAb and HRPA were added in turn with washing after each procedure, and then IgG4Ab-IgG4- Bio- IgG4Ab--HRP-A complex was formed on solid plate. After color display with enzyme substrate, OD value under 490 nm was measured and IgG4 concentration in the samples was calculated according to the standard curve. The detective range of this assay was from 2.5 to 200 ng/ mL with the lowest detection level of 2.1 ng/mL, and the intra-and inter-assay CV were 8.4% and 11.2%, respectively. Using this assay, the serum IgG4 level of normal youth detected was 37.7±2.3 ng/mL(n=30), that of blood-donator was 42.7±9.3ng/mL(n=30), that of patients in intensive care unit(ICU) was 71.2±9.3(n=49), which was significantly higher than that of normal youth, and that of patients with pulmonary infection was 28.4±9.2ng/mL(n= 49), which was significantly lower than that of normal youth. These results showed that this assay is reliable, and its sensitivity is appropriate for measuring serum IgG4 levels in normal individuals.
出处 《标记免疫分析与临床》 CAS 2006年第2期92-94,共3页 Labeled Immunoassays and Clinical Medicine
关键词 IGG4 生物素 亲和素 辣根过氧化物酶 IgG4 Biotin Avidin Horse radish peroxidase
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