Escherichia coli O157:H7 bacterium as a zoonotic pathogen is one of the most important causative agents of foodborne illnesses worldwide. Due to the serious concerns in public health and enormous economic losses in agriculture and food industry, it is very necessary to develop novel technology–based methods for sensitive and rapid detection of this bacterium in contaminated resources. In this study, a sensitive and selective fluorescence DNA nanosensing platform based on graphene oxide (GO) and the 3′ end of eae gene as specific sequence was developed for the detection of E. coli O157:H7. In this platform, fluorescence resonance energy transfer (FRET) process between GO- and FAM-labeled eae gene probe was used for the diagnosis of E. coli O157:H7. Following the immobilization of the eae gene probe on GO, fluorescence emission of FAM was quenched. In hybridization reaction, by adding the complementary DNA, fluorescence emission of FAM was significantly increased and recovered to 93%. The performance of sensor for detection of E. coli O157:H7 genomic DNA was determined 10 pg genomic DNA per 1 ml Tris-HCl hybridization buffer which was significantly more sensitive than PCR method. In conclusion, the results indicated that GO eae gene-based nanosensor has potential to be developed as a rapid and sensitive diagnostic device besides PCR methods for the detection of E. coli O157:H7 bacteria.

中文翻译：

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基于eae基因的荧光DNA纳米传感器3'端，用于检测大肠杆菌O157：H7。

大肠杆菌O157：H7细菌是一种人畜共患病原体，是全球食源性疾病最重要的致病因素之一。由于对公共卫生的严重关注以及农业和食品工业的巨大经济损失，因此有必要开发基于新技术的方法来灵敏，快速地检测受污染资源中的这种细菌。在这项研究中，开发了一种基于氧化石墨烯（GO）和eae基因3'末端作为特定序列的灵敏且选择性的荧光DNA纳米传感平台，用于检测大肠杆菌O157：H7。在这个平台上，GO和FAM标记的eae基因探针之间的荧光共振能量转移（FRET）过程被用于诊断糖尿病。大肠杆菌O157：H7。将eae基因探针固定在GO上后，FAM的荧光发射被淬灭。在杂交反应中，通过添加互补DNA，FAM的荧光发射显着增加并恢复到93％。每1 ml Tris-HCl杂交缓冲液测定10 pg基因组DNA时，检测大肠杆菌O157：H7基因组DNA的传感器性能显着高于PCR方法。总之，结果表明，基于GO eae基因的纳米传感器除用于检测大肠杆菌O157：H7细菌的PCR方法外，还有望发展成为一种快速灵敏的诊断设备。