摘要: AIM: To establish the rapid, specific, and sensitive method for detecting O157:H7 with DNA microchips. METHODS: Specific oligonucleotide probes (26-28 nt) of bacterial antigenic virulent genes E. coli other related pathogen were pre-synthesized immobilized on a solid support to make microchips. The four encoding O157 somatic antigen (rfbE), H7 flagellar (fliC) toxins (SLT1, SLT2) monitored by multiplex PCR pairs specific primers. Fluorescence-Cy3 labeled samples hybridization generated Cy3-labeled single prime. Hybridization was performed 60 min at 45 °C. Microchip images taken using confocal fluorescent scanner. RESULTS: Twelve different strains detected various combinations genes. All yielded positive results PCR. size products these primers varied from 210 678 bp. rfbE/fliC/SLT1/SLT2 specifically recognized strains, or containing No cross occurred in probes. Non-O157:H7 pathogens failed yield any signal under comparable conditions. If product diluted 50-fold, no found agarose gel electrophoresis, but microarray hybridization. CONCLUSION: Microarray analysis is efficient identification detection pathogens.
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