生物技术通报 ›› 2018, Vol. 34 ›› Issue (3): 75-79.doi: 10.13560/j.cnki.biotech.bull.1985.2017-0873

• 综述与专论 • 上一篇    下一篇

微囊藻毒素-LR的定量荧光纳米微球免疫层析检测

张艺1, 丁新良2, 张珏1, 周彬1, 郭明明1, 黄飚1   

  1. 江苏省原子医学研究所 卫生部核医学重点实验室 江苏省分子核医学重点实验室,无锡 214063;
    无锡市疾病预防控制中心,无锡 214023
  • 收稿日期:2017-10-19 出版日期:2018-03-20 发布日期:2018-04-10
  • 作者简介:张艺,女,硕士,研究方向生物医学;E-mailzhangyi@jsinm.org
  • 基金资助:
    国家自然科学(81502831),江苏省卫计委预防医学科研课题(Y2015072)

Quantitative Detection of Microcystin-LR by Fluorescent Nanosphere Immunochromatographic Assay

ZHANG Yi1, DING Xin-liang2, ZHANG Jue1, ZHOU Bin1, GUO Ming-ming1, HUANG Biao1,   

  1. 1. Key Laboratory of Nuclear Medicine,Ministry of Health & Jiangsu Key Laboratory of Molecular Nuclear Medicine,Jiangsu Institute of Nuclear Medicine,Wuxi 214063;
    2. Wuxi Center for Disease Control and Prevention,Wuxi 214023
  • Received:2017-10-19 Published:2018-03-20 Online:2018-04-10

摘要: 研制了一种快速、定量荧光免疫层析试纸条,用于检测水体中MC-LR含量。基于竞争法原理,采用荧光免疫层析技术,建立MC-LR免疫层析法,并对该试剂考察其灵敏度、准确度、特异性、稳定性等指标。经过对免疫层析试纸条制备参数的优化,即检测线抗原浓度、微球标记量和结合垫微球浓度的选择,加样15 min后,获得了MC-LR荧光免疫层析法的工作直线,方法灵敏度0.195 μg/L,回收率99.4%,变异系数小于10%,与MC-LF交叉反应率低,与市售试剂盒检测结果一致,试剂在37℃稳定保存6 d。本研究研制的荧光免疫层析试剂灵敏、稳定、准确,可快速、定量检测水样中MC-LR含量,操作便捷。

关键词: 免疫层析, 微囊藻毒素, 纳米微球, 荧光

Abstract: This study was designed to develop a rapid quantitative detection strip for MC-LR(microcystin-LR)in water samples using the fluorescent nanosphere immunochromatograhic technology. Based on competitive mode,an immunochromatograhic strip system was developed for detecting MC-LR,and the reagent performance was evaluated for its sensitivity,accuracy,specificity and stability. Several parameters for preparing immunochromatograhic strip,i.e.,the concentration of artificial antigen on the test line,labeling rate of nanosphere and antibody,the concentration of nanospheres on the conjugate pad,were optimized. After the sample added for 15 min,the immunochromatographic assay was finished with a linear working range,0.195 μg/L of sensitivity,99.4% of recovery,below 10% of variable coefficient,little cross reaction with MC-LF,well consistency with commercial kits,and solid stability for 6 days at 37℃. In summary,this fluorescent nanosphere immunochromatographic strip is sensitive,stable,accurate,rapid,quantitative,and easy performed for MC-LR detection.

Key words: immunochromatographic, microcystin, nanosphere, fluorescence