Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
2199922 | Molecular and Cellular Probes | 2010 | 6 Pages |
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.
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Authors
Yoshihiro Kouguchi, Takako Fujiwara, Miki Teramoto, Mika Kuramoto,