Detection of E. coli O157:H7 using its endogenous active membrane peroxidase

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Title
Detection of E. coli O157:H7 using its endogenous active membrane peroxidase
Author(s)
K H Shim; M Kang; M G Kim; Bong Hyun Chung; S S A An
Bibliographic Citation
Toxicology and Environmental Health Sciences, vol. 3, no. 2, pp. 80-85
Publication Year
2011
Abstract
Recently, new virulent strain of Escherichia coli (E. coli) bacteria (0104:H4) was identified, causing ongoing outbreak of food poisoning in Europe, infecting up to 1,600 people with 18 deaths, especially in Germany. More common strain of E. coli for the hemorrhagic colitis and hemolytic uremic syndrome in human from food poisoning was E. coli O157:H7 (O157). Hence, various diagnostic methods for detecting O157 using staining procedure, PCR after cell lysis, sandwich ELISA, electrochemical biosonsors, and SPR biosensor were developed to expedite the detection time. In this study, our aim was to detect and monitor O157 non-invasively without lysing cells by using its endogenous membrane peroxidase. The genomic analysis revealed that O157 had seven different peroxidases, however, their expression levels, locations, or activities had not been revealed. Various peroxidase substrates for the detection by absorption, fluorescence and luminescence spectroscopies revealed 3,3′,5,5′ Tetramethylbenzidine (TMB), as the best peroxidase substrate against O157. Using TMB, the membrane peroxidase of O157 could be detected in 30 to 60 min with the detection levels of 105 cells/mL, comparable to current ELISA kits. Since TMB is a well-known colorimetric HRP substrate without using sophisticated equipments and procedures, the quantification of O157 with the hand-held portable device could yield compatible detection limit. In future, in conjunction with conjugated anti-O157 antibody on magnetic beads, the present method of using endogenous membrane peroxidases could be further developed to a simple point of care test system to be used in the field.
Keyword
Endogeneous membrane peroxidaseEscherichia coliListeiraO157:H7Staphylococcus aureus
ISSN
2005-9752
Publisher
Springer
DOI
http://dx.doi.org/10.1007/s13530-011-0081-6
Type
Article
Appears in Collections:
1. Journal Articles > Journal Articles
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