Detection and identification of multiple genetically modified events using DNA insert fingerprinting.
Current screening and event-specific polymerase chain reaction (PCR) assays for the detection and identification of genetically modified organisms (GMOs) in samples of unknown composition or for the detection of non-regulated GMOs have limitations, and alternative approaches are required. A transgenic DNA fingerprinting methodology using restriction enzyme digestion, adaptor ligation, and nested PCR was developed where individual GMOs are distinguished by the characteristic fingerprint pattern of the fragments generated. The inter-laboratory reproducibility of the amplified fragment sizes using different capillary electrophoresis platforms was compared, and reproducible patterns were obtained with an average difference in fragment size of 2.4 bp. DNA insert fingerprints for 12 different maize events, including two maize hybrids and one soy event, were generated that reflected the composition of the transgenic DNA constructs. Once produced, the fingerprint profiles were added to a database which can be readily exchanged and shared between laboratories. This approach should facilitate the process of GMO identification and characterization.
Raymond
Philippe
P
St-Hyacinthe Laboratory, Canadian Food Inspection Agency (CFIA), St-Hyacinthe, QC J2S 8E3, Canada. philippe.raymond@inspection.gc.ca
Gendron
Louis
L
Khalf
Moustafa
M
Paul
Sylvianne
S
Dibley
Kim L
KL
Bhat
Somanath
S
Xie
Vicki R D
VR
Partis
Lina
L
Moreau
Marie-Eve
ME
Dollard
Cheryl
C
Coté
Marie-José
MJ
Laberge
Serge
S
Emslie
Kerry R
KR
D001483:Base Sequence; D016172:DNA Fingerprinting / Q000379:methods*; D018744:DNA, Plant / Q000737:chemistry / Q000235:genetics; D005818:Genetic Engineering; D008969:Molecular Sequence Data; D030821:Plants, Genetically Modified / Q000737:chemistry / Q000235:genetics*; D016133:Polymerase Chain Reaction; D016415:Sequence Alignment; D013025:Glycine max / Q000737:chemistry / Q000235:genetics*; D003313:Zea mays / Q000737:chemistry / Q000235:genetics*
D018744:DNA, Plant