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Rapid and Simple Colorimetric Detection of Escherichia coli O157:H7 in Apple Juice Using a Novel Recombinant Bacteriophage-Based Method

机译:基于新型噬菌体重组方法的苹果汁中大肠杆菌O157:H7的快速,简单比色检测

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摘要

In this study, a bacteriophage-based method for the colorimetric detection of E. coli O157:H7 in apple juice was investigated. Firstly, a gene encoding Cytochrome c Peroxidase (CCP) chromogenic enzyme was inserted into a wild type PP01 phage genome to construct the recombinant PP01ccp phage that was used in the production of the chromogenic enzyme through specific infection into E. coli O157:H7. The method was then examined in the colorimetric detection of E. call O157:H7 in broth, and the appearance of E. coil O157:H7 in broth was confirmed by the color change after a few minutes of the enzyme assay. Secondly, the method was investigated in the colorimetric detection of E. coli O157:H7 in apple juice. A low E. coil O157:H7 concentration as 1 CFU ml(-1) was detected in 15 h that was in a shorter time than in previous bioluminescence phage-based methods. Moreover, the method is much simpler compared to other previous phage-based methods since it enables detection without the need for expensive apparatus.
机译:在这项研究中,研究了一种基于噬菌体的比色法检测苹果汁中大肠杆菌O157:H7的方法。首先,将编码细胞色素c过氧化物酶(CCP)发色酶的基因插入野生型PP01噬菌体基因组,以构建重组PP01ccp噬菌体,该噬菌体用于通过特异性感染大肠杆菌O157:H7而产生发色酶。然后在肉汤中大肠杆菌O157:H7的比色检测中检查该方法,并通过酶分析几分钟后的颜色变化来确认肉汤中大肠杆菌O157:H7的出现。其次,对比色法检测苹果汁中大肠杆菌O157:H7的方法进行了研究。在15小时内检测到低的大肠杆菌O157:H7浓度为1 CFU ml(-1),这比以前的基于生物发光噬菌体的方法要短。此外,与其他先前的基于噬菌体的方法相比,该方法简单得多,因为它无需昂贵的设备即可进行检测。

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