ALF: a strategy for identification of unauthorized GMOs in complex mixtures by a GW-NGS method and dedicated bioinformatics analysis

Scientific Reports
Alexandra Bogožalec KoširJeroen P van Dijk

Abstract

The majority of feed products in industrialised countries contains materials derived from genetically modified organisms (GMOs). In parallel, the number of reports of unauthorised GMOs (UGMOs) is gradually increasing. There is a lack of specific detection methods for UGMOs, due to the absence of detailed sequence information and reference materials. In this research, an adapted genome walking approach was developed, called ALF: Amplification of Linearly-enriched Fragments. Coupling of ALF to NGS aims for simultaneous detection and identification of all GMOs, including UGMOs, in one sample, in a single analysis. The ALF approach was assessed on a mixture made of DNA extracts from four reference materials, in an uneven distribution, mimicking a real life situation. The complete insert and genomic flanking regions were known for three of the included GMO events, while for MON15985 only partial sequence information was available. Combined with a known organisation of elements, this GMO served as a model for a UGMO. We successfully identified sequences matching with this organisation of elements serving as proof of principle for ALF as new UGMO detection strategy. Additionally, this study provides a first outline of an automated, we...Continue Reading

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Citations

Mar 27, 2018·Analytical and Bioanalytical Chemistry·Tigst Demeke, David Dobnik
May 23, 2018·Scientific Reports·Marie-Alice FraitureNancy H C Roosens
Feb 28, 2020·Scientific Reports·Dong YuDingyang Yuan

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Datasets Mentioned

BETA
MON89034
MON15985
JQ406879
AY326434
US756989
GP69587
MON88017
EA135633

Methods Mentioned

BETA
PCRs
PCR
GMO
transgenic

Software Mentioned

PacBio RS II
Megablast
CFX Manager
PacBio SMRT
APAgene GOLD
BEDtools
BLAST
Bowtie2
excel
UPARSE

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