| 초록 |
DNA probe assay systems that can simultaneously analyze multiple genes at the places where specimen (e.g., food) is offered have been developed. Genes to be detected were prepared from food-contaminating microorganisms (E. coli O157:H7 and S. typhimurium in this study) and multiplex PCR (utilizing the reverse primers with a fluorescent label at each 5'-end) were then performed such that specific sequences of nucleic acids to each organism were amplified at the same time. These different DNA products from PCR were detected in a system consisting of capture probes, hybridizing with each PCR product, immobilized on spatially separated solid surfaces and signal generator with a catalytic action. The probe was synthesized to contain biotin at the 5'-end and bound onto either plastic or glass surfaces with immobilized streptavidin by employing ligand-receptor linkage. Signal-to-noise ratio yielded from the assay system was controlled by introducing an organic solvent (e.g., formamide) preventing non-specific interaction of excess primers that were present in the unpurified PCR mixture. We achieved without a significant sacrifice in the detection sensitivity, i.e., 1x105 CFU/mL for each gene. |
