Development of a peptide nucleic acid array platform for the detection of genetically modified organisms in food.
Two previously developed platforms, a multiplex polymerase chain reaction (PCR) and a peptide nucleic acid (PNA) array, the former allowing for the simultaneous detection of five transgenes and two endogenous controls in food and feed matrices and the latter for the assessment of the identity of amplified PCR products, were combined in order to develop a PNA array device for the screening of genetically modified organisms (GMOs) in food. PNA probes were opportunely designed, synthesized, and deposited on commercial slides. The length of the probes as well as the distance of the probes from the surface were evaluated and found to be critical points. The most suitable probes were found to be 15-mer PNAs linked to the slide surface by means of two 2-(2-aminoethoxy)ethoxyacetic acids as spacers. The device was tested on a model system constituted by flour samples containing a mixture of standards at known concentrations of transgenic material, in particular Roundup Ready soybean and Bt11, Bt176, Mon810, and GA21 maize: The DNA was amplified using the specific multiplex PCR method and tested on the PNA array. The method proposed was found to be able to correctly identify every GMO present in the tested samples.
Germini
Andrea
A
Dipartimento di Chimica Organica e Industriale, Università di Parma, Parco area delle Scienze 17/A, 43100 Parma, Italy.
Rossi
Stefano
S
Zanetti
Alessandro
A
Corradini
Roberto
R
Fogher
Corrado
C
Marchelli
Rosangela
R
D018744:DNA, Plant / Q000032:analysis*; D005504:Food Analysis / Q000379:methods*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D010455:Peptides / Q000235:genetics*; D030821:Plants, Genetically Modified / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods; D012680:Sensitivity and Specificity; D013025:Glycine max / Q000235:genetics; D019076:Transgenes / Q000235:genetics*; D003313:Zea mays / Q000235:genetics
D018744:DNA, Plant; D010455:Peptides