Amperometric DNA biosensor for the determination of auto-antibodies using DNA interaction with Pt(II) complex
Amperometric DNA biosensor for the determination of auto-antibodies using DNA interaction with Pt(II) complex
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DOI：
10.1016/j.aca.2003.09.057
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发表时间：
2004-01-23
影响因子：
6.2
通讯作者：
Zyavkina, YI
中科院分区：
文献类型：
--
作者：
Babkina, SS;Ulakhovich, NA;Zyavkina, YI
本文报道了一种变性DNA在硝酸纤维素膜上的固定方法。在固定汞膜覆盖银电极上制备了一种表面均匀稳定的修饰膜，并将其作为电流型DNA生物传感器的生物敏感元件。该生物传感器已被用于设计一种新的固相免疫分析血清中的自身抗体（Ab）的不分离的成分。通过测量由Pt（II）分别与DNA或自身抗体络合产生的催化析氢电流（电势为-1.2和-1.4V）来监测自身抗体的含量。在5.0 x 10（-10）至7.0 x 10（-8）M的宽浓度区域内进行测定。检测限为3.0 × 10 ~（-10）M。已发现DNA-抗体免疫反应的亲和常数为1.25 × 10（9）和2.50 × 10（8）M-1，这证实了相互作用的特异性。本文提出了水貂阿留申病（AMD）的免疫诊断方法，并对诊断步骤进行了描述。(C)2003 Elsevier B. V.保留所有权利。
A method of denatured DNA immobilisation on cellulose nitrate film has been developed. A modified film of uniform and stable surface has been used as a bio-sensitive part of amperometric DNA biosensor based on the stationary mercury-film covered silver electrode. The biosensor has been used to devise a new variant of solid-phase immumoassay of auto-antibodies (Ab) in blood serum without separation of components. The content of auto-Ab was monitored by measuring the currents of catalytic hydrogen evolution (with potentials of -1.2 and -1.4 V) resulting from the complexing of Pt(II) with DNA or auto-Ab respectively. The determination has been performed within a wide concentration area of 5.0 x 10(-10) to 7.0 x 10(-8) M. The limit of detection is 3.0 x 10(-10) M. The affinity constants for the immunoreaction of DNA-antibodies have been found to be 1.25 x 10(9) and 2.50 x 10(8) M-1, which confirms the specificity of the interaction. The protocol of the immunoassay has been proposed and the procedure of diagnosing Aleutian mink disease (AMD) has been described here. (C) 2003 Elsevier B.V. All rights reserved.