subtractive inhibition assay for the detection of e. coli o157h7 using surface plasmon resonance减去抑制试验的检测大肠杆菌o157h7使用表面等离子体共振.pdf

Sensors 2011, 11, 2728-2739; doi:10.3390/s110302728 OPEN ACCESS sensors ISSN 1424-8220 /journal/sensors Article Subtractive Inhibition Assay for the Detection of E. coli O157:H7 Using Surface Plasmon Resonance Yixian Wang, Zunzhong Ye *, Chengyan Si and Yibin Ying College of Biosystems Engineering and Food Science, Zhejiang University, Hangzhou 310029, Zhejiang, China; E-Mails:wang_yi_xian1986@ (Y.W.); sichengyanzhj@163.com (C.S.); ybying@ (Y.Y.) * Author to whom correspondence should be addressed; E-Mail: zzye@; Tel.: +86-571-8697-1885; Fax: +86-571-8697-1885. Received: 8 January 2011; in revised form: 29 January 2011 / Accepted: 21 February 2011/ Published: 1 March 2011 Abstract: A surface plasmon resonance (SPR) immunosensor was developed for the detection of E. coli O157:H7 by means of a new subtractive inhibition assay. In the subtractive inhibition assay, E. coli O157:H7 cells and goat polyclonal antibodies for E. coli O157:H7 were incubated for a short of time, and then the E. coli O157:H7 cells which bound antibodies were removed by a stepwise centrifugation process. The remaining free unbound antibodies were detected through interaction with rabbit anti-goat IgG polyclonal antibodies immobilized on the sensor chip using a BIAcore 3000 biosensor. The results showed that the signal was inversely correlated with the concentration of 4 8