Towards on-site testing of Phytophthora species

被引：21

作者：

Schwenkbier, Lydia ^{[1
,2
,3
,4
]}

Pollok, Sibyll ^{[1
,2
,3
,4
]}

Koenig, Stephan ^{[5
]}

Urban, Matthias ^{[1
]}

Werres, Sabine ^{[5
]}

Cialla-May, Dana ^{[1
,2
,3
,4
]}

Weber, Karina ^{[1
,2
,3
,4
]}

Popp, Juergen ^{[1
,2
,3
,4
]}

机构：

[1] Leibniz Inst Photon Technol Jena iPHT, Jenaer BioChip Initiat, D-07745 Jena, Germany

[2] Univ Jena, Inst Phys Chem, D-07745 Jena, Germany

[3] Univ Jena, Abbe Ctr Photon, D-07745 Jena, Germany

[4] InfectoGnost Forsch Campus Jena, Zentrum Angew Forsch, D-07743 Jena, Germany

[5] Julius Kuehn Inst, Fed Res Ctr Cultivated Plants JKI, Inst Plant Protect Hort & Forests, D-38104 Braunschweig, Germany

来源：

ANALYTICAL METHODS | 2015年 / 7卷 / 01期

关键词：

HELICASE-DEPENDENT AMPLIFICATION; MEDIATED ISOTHERMAL AMPLIFICATION; NUCLEIC-ACID AMPLIFICATION; MOLECULAR TOOL BOX; DNA EXTRACTION; STAPHYLOCOCCUS-AUREUS; ONCHIP-AMPLIFICATION; ASSAY; PATHOGEN; IDENTIFICATION;

D O I：

10.1039/c4ay02287d

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

Rapid detection and accurate identification of plant pathogens in the field is an ongoing challenge. In this study, we report for the first time on the development of a helicase-dependent isothermal amplification (HDA) in combination with on-chip hybridization for the detection of selected Phytophthora species. The HDA approach allows efficient amplification of the yeast GTP-binding protein (Ypt1) target gene region at one constant temperature in a miniaturized heating device. The assay's specificity was determined by on-chip DNA hybridization and subsequent silver nanoparticle deposition. The silver deposits serve as stable endpoint signals that enable the visual as well as the electrical readout. Our promising results point to the direction of a near future on-site application of the combined techniques for a reliable detection of Phytophthora species.

引用

页码：211 / 217

页数：7

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