Ravan Hadi, Amandadi Mojdeh, Sanadgol Nima
Department of Biology, Faculty of Science, Shahid Bahonar University of Kerman, Kerman, Iran.
Department of Biology, Faculty of Science, Shahid Bahonar University of Kerman, Kerman, Iran.
Microb Pathog. 2016 Feb;91:161-5. doi: 10.1016/j.micpath.2015.12.011. Epub 2015 Dec 25.
E. coli O157:H7 is one of the most important foodborne pathogen that causes some human illnesses such as bloody diarrhea, hemolytic-uremic syndrome, and kidney failure. We developed a loop-mediated isothermal amplification (LAMP) assay with six special primers that target a highly specific 299-bp region of the Z3276 gene for the detection of E. coli O157:H7. Among 117 bacterial strains tested in this study, positive results were only obtained from E. coli O157:H7 strains. The sensitivity level of the Z3276-LAMP assay was determined to be 5 CFU/reaction tube in pure bacterial culture. Moreover, the LAMP assay was successfully applied to artificially contaminated ground beef with a sensitivity level of 10(3) CFU/mL without pre-enrichment and 10 CFU/mL after a 4-h pre-enrichment. In conclusion, the present LAMP assay would be a useful and powerful tool for the rapid, sensitive, and specific diagnosis of E. coli O157:H7 strains in resource limited laboratories.
大肠杆菌O157:H7是最重要的食源性病原体之一，可导致一些人类疾病，如出血性腹泻、溶血尿毒综合征和肾衰竭。我们开发了一种环介导等温扩增（LAMP）检测方法，该方法使用六种特殊引物靶向Z3276基因的一个高度特异性的299碱基对区域，用于检测大肠杆菌O157:H7。在本研究中测试的117株细菌中，仅从大肠杆菌O157:H7菌株获得了阳性结果。Z3276-LAMP检测方法在纯细菌培养中的灵敏度水平确定为每反应管5 CFU。此外，LAMP检测方法成功应用于人工污染的绞碎牛肉，在不进行预富集时灵敏度水平为10³ CFU/mL，在4小时预富集后为10 CFU/mL。总之，目前的LAMP检测方法将是资源有限实验室中快速、灵敏和特异性诊断大肠杆菌O157:H7菌株的有用且强大的工具。
