Hairpin-DNA probe for enzyme-amplified electrochemical detection of Legionella pneumophila

被引：98

作者：

Miranda-Castro, Rebeca ^{[1
]}

de-los-Santos-Alvarez, Patricia ^{[1
]}

Lobo-Castanon, M. Jesus ^{[1
]}

Miranda-Ordieres, Arturo J. ^{[1
]}

Tunon-Blanco, Paulino ^{[1
]}

机构：

[1] Univ Oviedo, Dept Quim Fis & Analit, E-33006 Oviedo, Spain

来源：

ANALYTICAL CHEMISTRY | 2007年 / 79卷 / 11期

关键词：

D O I：

10.1021/ac062260q

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

An electrochemical genosensor for the detection of nucleic acid sequences specific of Legionella pneumophila is reported. An immobilized thiolated hairpin probe is combined with a sandwich-type hybridization assay, using biotin as a tracer in the signaling probe, and streptavidin-alkaline phosphatase as reporter molecule. The activity of the immobilized enzyme was voltammetrically determined by measuring the amount of 1-naphthol generated after 2 min of enzymatic dephosphorylation of 1-naphthyl phosphate. The sensor allows discrimination between L. pneumophila and L. longbeachae with high sensitivity under identical assay conditions (no changes in stringency). A limit of detection of 340 pM L. pneumophila DNA, and a linear relationship between the analytical signal and the logarithm of the target concentration to 2 mu M were obtained. Experimental results show the superior sensitivity and selectivity of the hairpin-based assay when compared with analogous sandwich-type assays using linear capture probes.

引用

收藏

页码：4050 / 4055

页数：6

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