犬C-反应蛋白荧光定量免疫层析检测方法的建立与应用

doi:10.16431/j.cnki.1671-7236.2019.02.034

《中国畜牧兽医》 ›› 2019, Vol. 46 ›› Issue (2): 621-627.doi: 10.16431/j.cnki.1671-7236.2019.02.034

犬C-反应蛋白荧光定量免疫层析检测方法的建立与应用

康业¹, 王羽¹, 胡树昆¹, 吴培钿², 李文美^1,2, 何小维¹

1. 华南理工大学食品科学与工程学院, 广州 510640;
2. 广州万孚生物技术股份有限公司, 广州 510663

收稿日期:2018-08-15 出版日期:2019-02-20 发布日期:2019-02-20
通讯作者: 何小维 E-mail:davidhxwhe@126.com
作者简介:康业(1994-),男,河南商丘人,硕士生,研究方向:POCT诊断试剂,E-mail:331491396@qq.com
基金资助:
广东省省级科技计划项目（2017A020208014）

Establishment and Application of Fluorescence Quantitive Immunochromatographic Assay for Canine C-reactive Protein

KANG Ye¹, WANG Yu¹, HU Shukun¹, WU Peidian², LI Wenmei^1,2, HE Xiaowei¹

1. College of Food Science and Engineering, South China University of Technology, Guangzhou 510640, China;
2. Guangzhou Wondfo Biotech Co., Ltd., Guangzhou 510663, China

Received:2018-08-15 Online:2019-02-20 Published:2019-02-20

摘要/Abstract

摘要：

试验开展了犬C-反应蛋白（C-CRP）荧光微球免疫层析定量方法的研究，旨在研制一种操作简易、灵敏度高，用于快速定量检测C-CRP荧光免疫层析试纸条。采用双抗体夹心法和荧光免疫层析技术，以羧基荧光微球标记的抗C-CRP单克隆抗体及羊抗鸡IgY为标记抗体，抗C-CRP单克隆抗体和鸡IgY分别作为检测线和质控线制备荧光免疫层析试纸条。结果显示，所制备C-CRP检测试纸条的检测范围为0.5~250 mg/L，检测限为0.5 mg/L，批内和批间变异系数（CV）分别为0.68%~6.94%和0.89%~8.79%，平均回收率为98.15%~101.19%，试剂与9种干扰物质无交叉反应。加速破坏稳定性试验表明试纸条可以常温放置24个月。临床样本测试发现，其与I-CHROMA试剂盒检测结果相关性良好（R²=0.995）。综上所述，该荧光免疫层析试纸条灵敏度、特异度较高，且操作简单、快速，可用于宠物犬临床检测。

关键词: 犬C-反应蛋白(C-CRP); 荧光免疫层析; 定量检测

Abstract:

The study focused on the establishment of fluorescent microsphere immunochromatography quantitative method of canine C-reactive protein (C-CRP) to a develop test strips of simple operation and high sensitivity for rapid quantitative detection of C-CRP.A fluorescent sandwich immunoassay via double antibody was set up,anti-C-CRP monoclonal antibody and goat anti-chicken IgY labeled with carboxy fluorescent microspheres were used as labeled antibody,the other anti-C-CRP monoclonal antibody and chicken IgY were used as capture antibody for detection lines and control line.The results showed that the detection range of the lateral flow assay was 0.5 to 250 mg/L,the detection limit was 0.5 mg/L,and the CV in intra-assay and inter-assay were 0.68% to 6.94% and 0.89% to 8.79%,respectively.The average recovery was 98.15% to 101.19%,and there was no cross-react with 9 interfering substances.The stressed condition stability test showed that the test strip could be stored for 24 months at room temperature.For clinical sample test,the test strips had a good correlation with the control reagent (R²=0.995).In conclusion,the prepared fluorescent immunochromatographic strips could be applied to clinical test of C-CRP concisely and conveniently.

Key words: canine C-reactive protein(C-CRP); fluorescence immunochromatography; quantitative detection

中图分类号:

S858.292

康业, 王羽, 胡树昆, 吴培钿, 李文美, 何小维. 犬C-反应蛋白荧光定量免疫层析检测方法的建立与应用[J]. 《中国畜牧兽医》, 2019, 46(2): 621-627.

KANG Ye, WANG Yu, HU Shukun, WU Peidian, LI Wenmei, HE Xiaowei. Establishment and Application of Fluorescence Quantitive Immunochromatographic Assay for Canine C-reactive Protein[J]. , 2019, 46(2): 621-627.

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犬C-反应蛋白荧光定量免疫层析检测方法的建立与应用

Establishment and Application of Fluorescence Quantitive Immunochromatographic Assay for Canine C-reactive Protein

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