Rapid and simple colorimetric detection of Escherichia coli O157:H7 in apple juice using a novel recombinant bacteriophage-based Method

Biocontrol Sci. 2015;20(2):99-103. doi: 10.4265/bio.20.99.

Abstract

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. coli O157:H7 in broth, and the appearance of E. coli 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. coli 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.

Publication types

  • Evaluation Study

MeSH terms

  • Bacteriological Techniques / methods*
  • Bacteriophages / genetics*
  • Bacteriophages / growth & development
  • Carbonated Beverages / microbiology*
  • Colorimetry / methods*
  • Cytochrome-c Peroxidase / analysis
  • Cytochrome-c Peroxidase / genetics
  • Escherichia coli O157 / isolation & purification*
  • Escherichia coli O157 / virology*
  • Food Microbiology / methods*
  • Genes, Reporter
  • Genome, Viral
  • Malus
  • Recombination, Genetic
  • Sensitivity and Specificity
  • Time Factors

Substances

  • Cytochrome-c Peroxidase