Hairpin-DNA Probe for Enzyme-Amplified Electrochemical Detection of Legionella pneumophila
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- 作者：Rebeca Miranda-Castro ; Patricia de-los-Santos-Á ; lvarez ; M. Jesú ; s Lobo-Castañ ; ó ; n ; Arturo J. Miranda-Ordieres ; Paulino Tuñ ; ó ; n-Blanco
- 刊名：Analytical Chemistry
- 出版年：2007
- 出版时间：June 1, 2007
- 年：2007
- 卷：79
- 期：11
- 页码：4050 - 4055
- 全文大小：169K
- 年卷期：v.79,no.11(June 1, 2007)
- ISSN：1520-6882
文摘
An electrochemical genosensor for the detection of nucleicacid sequences specific of Legionella pneumophila isreported. An immobilized thiolated hairpin probe iscombined with a sandwich-type hybridization assay, usingbiotin as a tracer in the signaling probe, and streptavidin-alkaline phosphatase as reporter molecule. The activityof the immobilized enzyme was voltammetrically determined by measuring the amount of 1-naphthol generatedafter 2 min of enzymatic dephosphorylation of 1-naphthylphosphate. The sensor allows discrimination between L.pneumophila and L. longbeachae with high sensitivityunder identical assay conditions (no changes in stringency). A limit of detection of 340 pM L. pneumophilaDNA, and a linear relationship between the analyticalsignal and the logarithm of the target concentration to 2
M were obtained. Experimental results show the superior sensitivity and selectivity of the hairpin-based assaywhen compared with analogous sandwich-type assaysusing linear capture probes.
M were obtained. Experimental results show the superior sensitivity and selectivity of the hairpin-based assaywhen compared with analogous sandwich-type assaysusing linear capture probes.