Detection of genetically modified organisms (GMOs) using isothermal amplification of target DNA sequences.
BACKGROUND
The most common method of GMO detection is based upon the amplification of GMO-specific DNA amplicons using the polymerase chain reaction (PCR). Here we have applied the loop-mediated isothermal amplification (LAMP) method to amplify GMO-related DNA sequences, 'internal' commonly-used motifs for controlling transgene expression and event-specific (plant-transgene) junctions.


RESULTS
We have tested the specificity and sensitivity of the technique for use in GMO studies. Results show that detection of 0.01% GMO in equivalent background DNA was possible and dilutions of template suggest that detection from single copies of the template may be possible using LAMP.


CONCLUSIONS
This work shows that GMO detection can be carried out using LAMP for routine screening as well as for specific events detection. Moreover, the sensitivity and ability to amplify targets, even with a high background of DNA, here demonstrated, highlights the advantages of this isothermal amplification when applied for GMO detection.
Lee
David
D
John Bingham Laboratory, National Institute of Agricultural Botany, Huntingdon Road, Cambridge, UK. david.lee@niab.com
La Mura
Maurizio
M
Allnutt
Theo R
TR
Powell
Wayne
W
D001483:Base Sequence; D029688:Brassica napus / Q000235:genetics; D017931:DNA Primers / Q000235:genetics; D018744:DNA, Plant / Q000235:genetics*; D008969:Molecular Sequence Data; D030821:Plants, Genetically Modified / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA / Q000379:methods; D019076:Transgenes
D017931:DNA Primers; D018744:DNA, Plant