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Rivista di Biologia Molecolare e Biotecnologie

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Minerva Biotecnologica 2004 September;16(3):203-10

lingua: Inglese

Real-­time detec­tion of genet­i­cal­ly mod­i­fied maize Bt-176 genom­ic sequenc­es by surface plasmon resonance-­based biospecific interaction analysis

Gardenghi S. 1, Finotti A. 1,2, Gambari R. 1, 2, Feriotto G. 1

1 Biotechnology Center, University of Ferrara, Ferrara, Italy;
2 Department of Biochemistry and Molecular Biology, University of Ferrara, Ferrara, Italy


The anal­y­sis of poly­me­rase-­chain reac­tion (PCR) prod­ucts by sur­face plas­mon res­o­nance (SPR) -based bio­spe­cif­ic inter­ac­tion anal­y­sis (BIA) ­using bio­sen­sors ­could be an excel­lent ­tool for spe­cif­ic and sen­si­tive detec­tion of genet­i­cal­ly mod­i­fied organ­isms (GMO). While ­this tech­nol­o­gy has sev­er­al impor­tant advan­tag­es, a ­major draw­back is ­that, in ­some instanc­es, sec­on­dary struc­tures of the PCR prod­ucts to be ana­lysed ­lead to low effi­cien­cy in bind­ing to the spe­cif­ic ­probes. Cross-hybrid­iza­tion ­between PCR prod­ucts and ­probes ­could ­also ­occur, espe­cial­ly ­when stan­dard BIA con­di­tions are ­employed. Here we dem­on­strate ­that bio­spe­cif­ic inter­ac­tion anal­y­sis, employ­ing sur­face plas­mon res­o­nance and bio­sen­sor tech­nol­o­gies, is suit­able to ­detect Bt-176 “maximizer” sequenc­es in ­maize. Not all the ­probes and PCR prod­ucts ­were ­found to be suit­able for SPR-­based BIA. However, we ­designed and char­ac­ter­ized ­probes ­able to selec­tive­ly iden­ti­fy and dis­crim­i­nate Bt-176 and ­zein ­gene sequenc­es exhib­it­ing ­high hybrid­isa­tion effi­cien­cy and no ­cross-hybrid­iza­tion. The ­results ­obtained sug­gest ­that SPR-­based BIA is a flex­ible, ­easy, ­speedy and auto­mat­able ­approach to ­detect genet­i­cal­ly mod­i­fied Bt-176 ­maize in ­foods.

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