Immobilization of Escherichia coli for detection of phage T4 using surface plasmon resonance |
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Authors: | Xiao ChangQing Jiang FengLei Zhou Bo Li Ran Liu Yi |
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Institution: | Key Laboratory of Virology;Key Laboratory of Analytical Chemistry for Biology and Medicine(Ministry of Edueation);College of Chemistry and Molecular Sciences,Wuhan University,Wuhan 430072,China 2 Department of Chemistry and Life Science,Hubei University of Education,Wuhan 430205,China 3 Department of Chemistry and Life Sciences,Xianning University,Xianning 437005,China |
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Abstract: | 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 1×10 7 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. |
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Keywords: | phage detection virus detection surface plasmon resonance(SPR) microbe-based biosensor self-assembled monolayer(SAM) |
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