氧化型α1抗胰蛋白酶定量检测试剂盒研制及对吸烟男性的检测分析

基础医学与临床 ›› 2013, Vol. 33 ›› Issue (6): 731-735.

氧化型α1抗胰蛋白酶定量检测试剂盒研制及对吸烟男性的检测分析

李振军¹,樊一笋²,Sam Alam³,陈菲⁴

1. 上海交通大学附属苏州九龙医院特需医疗科
2. 上海交通大学苏州九龙医院
3. Department of Medicine, Cambridge University
4. 苏州和锐医药科技有限公司

收稿日期:2013-02-18 修回日期:2013-04-25 出版日期:2013-06-05 发布日期:2013-05-29
通讯作者: 陈菲 E-mail:cf@herui.co

Introduction to oxidized α1-antitrypsin quantitative detection kit and its use in testing Chinese male smokers

Received:2013-02-18 Revised:2013-04-25 Online:2013-06-05 Published:2013-05-29

摘要/Abstract

摘要： 目的介绍一种血清氧化型α1抗胰蛋白酶（Ox-AT）的定量检测方法，并分析吸烟男性血清Ox-AT的表达水平。方法构建一种以Ox-AT特异性单抗为检测抗体的双抗夹心ELISA法，用纯化的Ox-AT作为标准品定量；分析我国吸烟与非吸烟男性血清中Ox-AT的水平。结果所建立的双抗夹心ELISA试剂盒检测限为2.58 μg/L，在0～500μg/L浓度范围内R2值0.9955，批间差为11.27%，准确度为88.19%，试剂盒4℃保质期超过6个月；对20对吸烟与非吸烟男性血清Ox-AT进行检测，吸烟组为55.43 ± 49.94 μg/L, 显著高于非吸烟对照组的11.01 ± 12.15 μg/L (p<0.001)，两组间α1-AT无显著差异。结论所建立的ELISA法能用于血清Ox-AT的定量检测；血清Ox-AT可能比α1-AT更适合作为我国COPD的评价指标。

关键词: 氧化型a1抗胰蛋白酶, 酶联免疫吸附法, 慢性阻塞性肺气肿

Abstract: Objective To introduce a serum oxidized α1-antitrypsin (Ox-AT) quantitative detection kit, and analyze serum Ox-AT in Chinese male smokers. Method Set up a sandwich ELISA method with Ox-AT and a specific anti-Ox-AT monoclonal antibody. Analyze serum Ox-AT level of 20 pairs of smoking and non-smoking male. Results Ox-AT sELISA has a good linear in range of 0-500μg/L. The limit detection is 2.58μg/L, CV among batches is 11.27%, the accuracy is about 88.19%, the shelf life under 4℃ is more than 6 months. Detecting with this sELISA, serum Ox-AT level in smoking male is 55.43 ± 49.94μg/L, non-smoking control group is 11.01 ± 12.15μg/L, p <0.001. Conclusion The Ox-AT detection kit can be used for quantitative testing serum Ox-AT. Ox-AT might be more useful than α1-AT in evaluating Chinese COPD.

Key words: α1-antitrypsin, ELISA, Chronic obstructive pulmonary disease

李振军樊一笋 Sam Alam 陈菲. 氧化型α1抗胰蛋白酶定量检测试剂盒研制及对吸烟男性的检测分析[J]. 基础医学与临床, 2013, 33(6): 731-735.