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基于使用未经处理抗血清的多重高灵敏度免疫层析测定法。

Multiplex highly sensitive immunochromatographic assay based on the use of nonprocessed antisera.

作者信息

Byzova Nadezhda A, Urusov Alexandr E, Zherdev Anatoly V, Dzantiev Boris B

机构信息

A.N. Bach Institute of Biochemistry, Federal Research Center "Fundamentals of Biotechnology", Leninsky prospeсt 33-2, 119071, Moscow, Russia.

出版信息

Anal Bioanal Chem. 2018 Mar;410(7):1903-1910. doi: 10.1007/s00216-018-0853-9. Epub 2018 Jan 20.

Abstract

The format of an immunochromatographic multiassay is first proposed with native antisera and a universal conjugate of antispecies antibodies with gold nanoparticles. This format allows (1) the exclusion of purification and conjugation stages for specific antibodies and (2) significant reduction of the concentration of specific antibodies in the system. The independent use of specific antibodies and a conjugated marker provided a low detection limit and high signal intensity. The proposed format was implemented for the simultaneous detection of two herbicides. The instrumental limits for the detection of atrazine and chlorsulfuron were 0.1 and 0.7 ng/mL, respectively, and the analysis time was 20 min. The suitability of the test system for monitoring these herbicides in nontreated apple and blackcurrant juices is shown. The assay technique is simple, sensitive, and easily transferrable to any other antigen. Graphical abstract The proposed format of the immunochromatographic multiassay is based on the use of native antisera and a universal conjugate of antispecies antibodies with gold nanoparticles. In this way purification and conjugation stages for specific antibodies are excluded, and the concentrations of specific antibodies and the conjugated marker can be varied independently to obtain a low detection limit.

摘要

免疫层析多重检测法的形式最初是用天然抗血清和抗种属抗体与金纳米颗粒的通用缀合物提出的。这种形式允许:(1)排除特异性抗体的纯化和缀合步骤;(2)显著降低系统中特异性抗体的浓度。特异性抗体和缀合标记物的独立使用提供了低检测限和高信号强度。所提出的形式被用于同时检测两种除草剂。检测莠去津和氯磺隆的仪器检测限分别为0.1和0.7 ng/mL,分析时间为20分钟。展示了该测试系统用于监测未处理苹果汁和黑加仑汁中这些除草剂的适用性。该检测技术简单、灵敏,并且易于转移到任何其他抗原上。图形摘要:所提出的免疫层析多重检测法的形式基于天然抗血清和抗种属抗体与金纳米颗粒的通用缀合物的使用。通过这种方式,排除了特异性抗体的纯化和缀合步骤,并且可以独立改变特异性抗体和缀合标记物的浓度以获得低检测限。

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