高等学校化学学报 ›› 2018, Vol. 39 ›› Issue (1): 41-47.doi: 10.7503/cjcu20170494

• 分析化学 • 上一篇    下一篇

背景荧光猝灭-免疫层析法检测C-反应蛋白

陈俊蕾1, 张唯2, 李新霞2, 李久彤3, 关明1()   

  1. 1. 新疆师范大学化学化工学院, 乌鲁木齐 830054
    2. 新疆医科大学药学院, 乌鲁木齐 830011
    3. 上海鑫谱生物科技有限公司, 上海 201800
  • 收稿日期:2017-07-21 出版日期:2018-01-10 发布日期:2017-12-04
  • 作者简介:联系人简介: 关 明, 男, 博士, 教授, 主要从事仪器分析新方法的研究. E-mail:guanm@xjnu.edu.cn
  • 基金资助:
    国家自然科学基金(批准号: 21465023)和新疆师范大学化学重点学科项目资助

Detection of C-reactive Protein by Background Fluorescence Quenching-Immunochromatography

CHEN Junlei1, ZHANG Wei2, LI Xinxia2, LI Jiutong3, GUAN Ming1,*()   

  1. 1. College of Chemistry and Chemical Engineering, Xinjiang Normal University, Urumqi 830054, China
    2. School of Pharmacy, Xinjiang Medical University, Urumqi 830011, China
    3. Shanghai Simp Bio-science Co., Ltd., Shanghai 201800, China
  • Received:2017-07-21 Online:2018-01-10 Published:2017-12-04
  • Contact: GUAN Ming E-mail:guanm@xjnu.edu.cn
  • Supported by:
    † Supported by the National Natural Science Foundation of China(No.21465023) and the Chemistry Key Discipline Project of Xinjiang Normal University, China

摘要:

采用背景荧光猝灭-免疫层析法(bFQICA), 基于双抗体夹心法原理, 将合适浓度的捕获抗体、 示踪抗体分别固定在试纸条和微孔内, 层析时与样本形成抗体-抗原-抗体的夹心结构, 建立了一种操作简便、 灵敏度高、 抗干扰性强且能快速定量检测C-反应蛋白(CRP)的方法. 结果表明, CRP在0.0~100.0 ng/mL浓度范围内与相对荧光强度值(F1/F2)呈现良好的相关性, 最低检出限为0.0939 ng/mL, 加标回收率为87.69%~111.0%, 检测3批试剂的批间和批内相对标准偏差均小于15%, 与降钙素原(PCT, 20.0 ng/mL)及人血清淀粉样蛋白A(SAA,10.0 μg/mL)均无交叉反应. 采用该方法与免疫透射比浊法同时测定41例临床血清样本, 检测结果相关性良好(r=0.9585, P<0.01), 2种方法无显著性差异(P>0.05).

关键词: 背景荧光猝灭-免疫层析法, C-反应蛋白, 胶体金

Abstract: Background

fluorescence immunochromatography assay(bFQICA) has been established, which has the advantages of simple operation, high sensitivity, strong anti-interference and rapid quantitative detection of C-reactive protein(CRP). This method of using double antibody sandwich method principle, the appropriate concentration of capture antibody and tracer antibodies are respectively fixed on the test strip and the micropore. During chromatography, antibodies were captured, while tracer antibodies and samples were inserted to form a sandwich structure of antibody antigen antibody. The method has a good correlation with fluorescence signal values(F1/F2) in the range of 0.0—100.0 ng/mL concentration in CRP, the minimum detection limit was 0.0939 ng/mL, and the recovery rate was 87.69%—111.0%, three batches of reagents and inter and intra assay relative standard deviation was less than 15%, and procalcitonin(PCT, 20.0 ng/mL), serum amyloid A like protein(SAA, 10.0 μg/mL) had not cross reaction with this method and the immune turbidimetric method for the simultaneous determination of 41 serum samples, the detection results had a good correlation(r=0.9585, P<0.01), there was no significant difference between the two methods(P>0.05). This method has high sensitivity, simple operation, and can be used for rapid and accurate quantitative detection of CRP.

Key words: Background fluorescence quenching-immunochromatography, C-Reactive protein, Colloidal gold