大肠杆菌O157菌体抗原基因检测芯片的制备

Detection of Escherichia coli O157 antigen gene with DNA microchips

选择编码O157菌体抗原特异合成酶的rfbE基因设计引物和探针,并制备检测芯片,通过两次PCR扩增,制备荧光标记的靶序列,并与芯片进行杂交,检测O157菌株和非O157病原体。结果所有O157菌株均在芯片相应探针处出现阳性信号,非O157杂交结果均为阴性;芯片检测灵敏度比PCR检测高50倍。说明基因芯片可以快速、灵敏、特异地检测O157菌体抗原,为建立快速灵敏的检测细菌病原体特征和鉴别诊断的自动分析系统提供了新方法。

Rapid, specific and sensitive DNA microchips are used for detection of O157 antigen gene. Primers and specific probes were designed from 0 antigen biosynthesis gene rfbE. The fluorescent-labeled PCR products were prepared by incorporation of Cy3-dUTP or Cy3-labeled primer during PCR amplification and were hybridized to the microchips for detection of O157 strains and non O157 pathogens. Positive signal were displayed on the mycrchips in all O157 strains, but non O157 pathogens failed to yield any signal under comparable conditions. The sensitivity of detection by microchips was shown to be at least 50 times than conventional PCR amplification. This mycroarray analysis are more specifically、rapidly、efficient than traditional bacterial detection. It might be very useful for automated identification and characterization of bacterial pathogens