Homogenous, real-time duplex loop-mediated isothermal amplification using a single fluorophore-labeled primer and an intercalator dye: Its application to the simultaneous detection of Shiga toxin genes 1 and 2 in Shiga toxigenic Escherichia coli isolates

被引：41

作者：

Kouguchi, Yoshihiro ^{[1
]}

Fujiwara, Takako ^{[1
]}

Teramoto, Miki ^{[1
]}

Kuramoto, Mika ^{[1
]}

机构：

[1] Fukuyama City Publ Hlth Ctr, Hiroshima, Japan

来源：

MOLECULAR AND CELLULAR PROBES | 2010年 / 24卷 / 04期

关键词：

Ethidium bromide; LAMP; Multiplex detection; Simultaneous detection; DISCRIMINATION; ASSAY; DNA; PCR;

D O I：

10.1016/j.mcp.2010.03.001

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

We developed a completely homogeneous duplex loop-mediated isothermal amplification (LAMP) method. The present LAMP method employed a combination of a 6-carboxyfluorescein (FAM)-labeled primer (donor) for one target gene, a non-labeled primer for the other, and an intercalator ethidium bromide (EtBr) dye (acceptor) on the basis of fluorescence resonance energy transfer (FRET) between the FAM donor and EtBr acceptor. Measuring changes in fluorescence of FAM enabled the LAMP method to detect two different genes simultaneously. This method was used to detect Shiga toxin genes in Shiga toxigenic Escherichia coli isolates, demonstrating simultaneous detection of two different genes with rapidity and accuracy. (C) 2010 Elsevier Ltd. All rights reserved.

引用

收藏

页码：190 / 195

页数：6

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