Zhang Meiling, Wang Xiudan, Han Lingzhi, Niu Shuyan, Shi Chao, Ma Cuiping
Key Laboratory of Sensor Analysis of Tumor Marker, Ministry of Education, College of Chemistry and Molecular Engineering, Qingdao University of Science and Technology, Qingdao 266042, PR China.
College of Marine Science and Biological Engineering, Qingdao University of Science and Technology, Qingdao 266042, PR China.
Anal Biochem. 2018 Mar 15;545:38-42. doi: 10.1016/j.ab.2018.01.013. Epub 2018 Jan 24.
A strand exchange amplification (SEA) method to detect foodborne pathogen Listeria monocytogenes was developed. SEA is a novel nucleic acid amplification method that only requires one pair of primers. The specie-specific primers were designed by targeting the 16S rRNA gene and the amplification reaction was performed as short as 60 min at 61 °C. Notably, SEA method could not only detect genomic DNA but also detect RNA by one step without requiring extra reverse transcription. The result could be visualized by naked eyes so that water bath pot would be the only equipment needed. Moreover, culture fluids and bacteria colony could be successfully detected without any pretreatment and the method displayed good specificity and strong anti-jamming capacity. These features greatly simplified the operating procedure and made SEA method be potential for developing point-of-care testing (POCT) devices to detect viable L. monocytogenes.
开发了一种用于检测食源性病原体单核细胞增生李斯特菌的链置换扩增(SEA)方法。SEA是一种新型核酸扩增方法,仅需一对引物。通过靶向16S rRNA基因设计种特异性引物,扩增反应在61°C下只需60分钟即可完成。值得注意的是,SEA方法不仅可以检测基因组DNA,还可以一步检测RNA,无需额外的逆转录。结果可用肉眼观察,因此水浴锅将是唯一所需的设备。此外,无需任何预处理即可成功检测培养液和菌落,该方法具有良好的特异性和较强的抗干扰能力。这些特点极大地简化了操作程序,使SEA方法具有开发即时检测(POCT)设备以检测活的单核细胞增生李斯特菌的潜力。