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	微孔板化学发光酶免疫分析法分析人血清中游离前列腺特异性抗原
	石根 ; 唐宝军 ; 王栩 ; 赵利霞 ; 林金明 ; Shi Gen ; Tang Bao-Jun ; Wang Xu ; Zhao Li-Xia ; Lin Jin-Ming
	2010-06-10 ; 2010-06-10
关键词	游离前列腺特异抗原 前列腺癌 化学发光酶免疫分析 肿瘤标志物 Free-prostate-specific antigen, prostate cancer, chemiluminescence enzyme immunoassay, tumor marker R446.6
其他题名	Microplate Chemiluminescence Enzyme Immunoassay for the Quantitative Analysis of Free Prostate-specific Antigen in Human Serum
中文摘要	建立了一种定量分析人血清中游离前列腺特异性抗原(f-PSA)的高灵敏度微孔板化学发光酶免疫分析方法,以碱性磷酸酶(alkaline phosphatase,ALP)为标记酶,4-甲氧基-4-(3″-磷酰氧基苯)-螺旋-(1,2-二氧杂环丁烷-3,2′-金刚烷)(4-methoxy-4-(3″-phosphate-phenyl)-spiro-(1,2-dioxetane-3,2′-adamantane),AMPPD)-ALP高灵敏的化学发光反应为检测体系,通过检测发光强度对人血清中f-PSA进行定量。对几种物理化学参数如温育时间、免疫反应步骤以及检测时间等进行了优化,采用了一步双抗体夹心法,37℃恒温静置温育2h,洗涤后加入50μL AMPPD,30~80min内检测。该方法线性范围为0.15~20μg/L,相关系数大于0.998;检出限可达0.01μg/L,批内和批间相对标准偏差(C.V.)均小于7%;回收率在88%~108%之间。对人体血清中共存的6种常见肿瘤标志物CA50、CA125、CA15-3、CA242、CEA和AFP的偶联反应进行考察,特异性良好。为了验证该方法用于商业试剂盒的可行性,在4℃和37℃条件下分别进行了3d,5d,7d的稳定性考察,其线性相关系数仍均大于0.998,相对标准偏差小于6%。对98例实际血样进行测定,并与进口试剂盒(Monobind.USA)作临床比对,相关性良好。这些结果均表明,该分析体系稳定,可靠,可以用于商业化试剂盒的开发,在临床分析人血清中f-PSA的辅助诊断前列腺癌方面具有很高的应用价值。; A microplate chemiluminescence enzyme immonoassay with high sensitivity for the quantitative analysis of free prostate specific antigen (f-PSA) in human serum was established. The immunoassay utilized alkaline phosphatase (ALP) as the labeled enzyme, and the highly sensitive ALP-AMPPD (4-methoxy-4-(3″-phosphate-phenyl)-spiro-(1,2-dioxetane-3,2′-adamantane)) chemiluminescence reaction as the detection system. Several physicochemical parameters such as incubation time, immunoreaction sequence, and detection time were studied and optimized. A one-step sandwiched reaction mode was applied with incubation at 37℃ for 2 h and chemiluminescent detection with 50 μL AMPPD for 30-80 min. The linear range of the proposed method for f-PSA was 0.15-20 μg/L with r=0.998. The detection limit was 0.01 μg/L. Inter-assay and intra-assay RSD were both below 7%. The recovery was 88%-108%. The cross-reactivity of six normal tumor markers in human serum, including CA50, CA125, CA15-3, CA242, CEA and AFP, was studied. It was shown that this method exhibited high specificity to f-PSA. The stability of the method was evaluated by identifying its analytical performance after stored at 4℃ and 37℃ for 3, 5, 7 days with RSD lower than 6% and correlation coefficient higher than 0.998, which showed satisfied reliability and stability. Compared with CLIA kit purchased from Monobind. Inc. USA, there was a good correlation between the two methods. The proposed method has shown great potential in the clinical analysis of f-PSA for prostate cancer diagnosis, as well as in the commercial kits development.; 国家自然科学基金资助项目(No20621703); 中国科学院人事局人才引进项目的支持
语种	中文 ; 中文
内容类型	期刊论文
源URL	[http://hdl.handle.net/123456789/60910]
专题	清华大学
推荐引用方式 GB/T 7714	石根,唐宝军,王栩,等. 微孔板化学发光酶免疫分析法分析人血清中游离前列腺特异性抗原[J],2010, 2010.
APA	石根.,唐宝军.,王栩.,赵利霞.,林金明.,...&Lin Jin-Ming.(2010).微孔板化学发光酶免疫分析法分析人血清中游离前列腺特异性抗原..
MLA	石根,et al."微孔板化学发光酶免疫分析法分析人血清中游离前列腺特异性抗原".(2010).

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