摘要
目的设计和制备快速、特异、灵敏的检测大肠埃希菌O157∶H7基因芯片。方法选择O157∶H7特异的rfbE、fliC、SLT1和SLT2基因,设计引物和探针,并制备检测芯片,通过两次聚合酶链反应(PCR)扩增,制备荧光标记的靶序列,并与芯片进行杂交,检测O157∶H7菌株和非O157病原体。结果O157∶H7菌株在采用单一和多重PCR两种方法制备的荧光标记靶序列与芯片杂交,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。结论基因芯片可以快速、灵敏、特异地检测O157∶H7,为建立快速灵敏的检测细菌病原体和鉴别诊断的自动分析系统提供了新方法。
Objective Rapid、specific and sensitive DNA microchips are used for detection of O157∶H7. Methods Primers and specific probes were designed from O157∶H7 gene rfbE、fliC、SLT1和SLT2. The fluorescent-labeled PCR products were prepared by incorporation of Cy3-labeled primer during PCR amplification and were hybridized to the microchips for detection of O157∶H7 strains and non O157 pathogens. Results Positive signal were all displayed on the mycrochips in O157∶H7 strains when using conventional PCR or multiplex PCR, but non O157 pathogens failed to yield any signal under comparable conditions. The sensitivity of detection by microchips was shown higher than PCR amplification. Conclusions This mycroarray analysis are more specifically、rapidly、efficient than traditional bacterial detection. It might be very useful for automated identification and detection of bacterial pathogens.
出处
《中华传染病杂志》
CAS
CSCD
北大核心
2005年第2期79-82,共4页
Chinese Journal of Infectious Diseases
基金
全军医学科学技术"十五"计划资助项目(01L006)
