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.

Abstract:

: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.

journal_name

Mol Cell Probes

authors

Kouguchi Y,Fujiwara T,Teramoto M,Kuramoto M

doi

10.1016/j.mcp.2010.03.001

subject

Has Abstract

pub_date

2010-08-01 00:00:00

pages

190-5

issue

4

eissn

0890-8508

issn

1096-1194

pii

S0890-8508(10)00014-9

journal_volume

24

pub_type

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