Immobilization of Escherichia coli for detection of phage T4 using surface plasmon resonance
Phage contamination is a very serious and unavoidable problem in modern fermentation industry. It is necessary to develop sensitive and rapid phage detection methods for the early detection of phage contamination. In the present work, a real-time, rapid, specific and quantitative phage T4 detection method based on surface plasmon resonance (SPR) technique has been introduced. Escherichia coli was immobilized onto the preformed MPA self-assembled monolayer (SAM) through the widely used EDC/NHS cross-linking reaction as the recognition element. The bacteria immobilization was verified efficiently through the electrochemical measurements and fluorescence microscopy observations. The specific adsorption was much stronger than the non-specific adsorption of phage T4 binding to the biosensor surface modified by E. coli, and the latter could be neglected. The detection sensitivity reached 1×107 PFU/mL within 10 min. Within the experimental phage concentrations, the linear correlation between the SPR response and the phage concentration was good. The results suggest that the SPR technique is a potentially powerful tool for the phage or other virus detections, as a label-free, real-time, and rapid method.
Keywordsphage detection virus detection surface plasmon resonance (SPR) microbe-based biosensor self-assembled monolayer (SAM)
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- 1.Duckworth DH. Who discovered bacteriophage? Bacteriol Rev, 1976, 40: 793–802Google Scholar
- 4.Jones DT, Shirley M, Wu X, Keis S. Bacteriophage infections in the industrial acetone butanol (AB) fermentation process. J Mol Microbiol Biotechnol, 2000, 2: 21–26Google Scholar
- 5.Łoś M, Czyż A, Sell E, Węgrzyn A, Neubauer P, Węcgrzyn G. Bacteriophage contamination: Is there a simple method to reduce its deleterious effects in laboratory cultures and biotechnological factories? J Appl Genet, 2004, 45: 111–120Google Scholar
- 16.Sambrook J, Russell DW. Molecular Cloning: A Laboratory Manual. 3rd ed. New York: Cold Spring Harbor Laboratory Press, 2001Google Scholar
- 20.Song F, Zhou F, Wang J, Tao N, Lin J, Vellanoweth RL, Morquecho Y, Laidman JW. Detection of oligonucleotide hybridization at femtomolar level and sequence-specific gene analysis of the Arabidopsis thaliana leaf extract with an ultrasensitive surface plasmon resonance spectrometer. Nucleic Acids Res, 2002, 30: e72CrossRefGoogle Scholar
- 22.Nagata K, Handa H. Real-time Analysis of Biomolecular Interactions: Applications of BIACORE. Berlin: Springer Verlag, 2000Google Scholar
- 26.Furukawa H, Yamada H, Mizushima S. Interaction of bacteriophage T4 with reconstituted cell envelopes of Escherichia coli K-12. J Bacteriol, 1979, 140: 1071–1080Google Scholar
- 30.Yu F, Mizushima S. Roles of lipopolysaccharide and outer membrane protein OmpC of Escherichia coli K-12 in the receptor function for bacteriophage T4. J Bacteriol, 1982, 151: 718–722Google Scholar