DETECTION OF GENETICALLY MODIFIED COMPONENTS BY A NEW RELIABLEMETHOD-CAPTURE-PCR-ELISA

摘要

Capture primer, the 5' terminus phosphorylated oligonucleotides, was covalently bound to the activated surface of PCR tubes. The target DNA fragment of the genetically modified soybean are hybridized with capture primer and other non-target elements in the sample preparation are removed by washing. The captured target DNAs were asymmetrically amplified and the amplicons are covalently immobilized on the surface of the PCR tubes. Immobilized amplicons were denatured, and hybridized with biotin-labeledoligo-probe and then detected by streptavidin-alkaline phosphatase. The method was specific and used to identify the GMO products in the international transportation. Parameters for binding of target DNA to the PCR tube and for PCR-ELISA detection of the covalently bound amplicon had been optimized. The sensitivity of the method was 10-100 DNA copy of templates/ul.