Subtractive Inhibition Assay for the Detection of E. coli O157:H7 Using Surface Plasmon Resonance
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...
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MDPI AG
2011-03-01
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| Series: | Sensors |
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| Online Access: | http://www.mdpi.com/1424-8220/11/3/2728/ |
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| author | Chengyan Si Yibin Ying Yixian Wang Zunzhong Ye |
| author_facet | Chengyan Si Yibin Ying Yixian Wang Zunzhong Ye |
| author_sort | Chengyan Si |
| collection | DOAJ |
| description | 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 E. coli O157:H7 cells in a range from 3.0 × 104 to 3.0 × 108 cfu/mL with a detection limit of 3.0 × 104 cfu/mL. Compared with direct SPR by immobilizing antibodies on the chip surface to capture the bacterial cells and ELISA for E. coli O157:H7 (detection limit: both 3.0 × 105 cfu/mL in this paper), the detection limit of subtractive inhibition assay method was reduced by one order of magnitude. The method simplifies bacterial cell detection to protein-protein interaction, which has the potential for providing a practical alternative for the monitoring of E. coli O157:H7 and other pathogens. |
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| issn | 1424-8220 |
| language | English |
| last_indexed | 2024-04-13T08:23:03Z |
| publishDate | 2011-03-01 |
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| spelling | doaj.art-02089fb4dc534a2e82eb8bd8a68342da2022-12-22T02:54:34ZengMDPI AGSensors1424-82202011-03-011132728273910.3390/s110302728Subtractive Inhibition Assay for the Detection of E. coli O157:H7 Using Surface Plasmon ResonanceChengyan SiYibin YingYixian WangZunzhong YeA 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 E. coli O157:H7 cells in a range from 3.0 × 104 to 3.0 × 108 cfu/mL with a detection limit of 3.0 × 104 cfu/mL. Compared with direct SPR by immobilizing antibodies on the chip surface to capture the bacterial cells and ELISA for E. coli O157:H7 (detection limit: both 3.0 × 105 cfu/mL in this paper), the detection limit of subtractive inhibition assay method was reduced by one order of magnitude. The method simplifies bacterial cell detection to protein-protein interaction, which has the potential for providing a practical alternative for the monitoring of E. coli O157:H7 and other pathogens.http://www.mdpi.com/1424-8220/11/3/2728/subtractive inhibition assaySPRE. coli O157:H7 |
| spellingShingle | Chengyan Si Yibin Ying Yixian Wang Zunzhong Ye Subtractive Inhibition Assay for the Detection of E. coli O157:H7 Using Surface Plasmon Resonance Sensors subtractive inhibition assay SPR E. coli O157:H7 |
| title | Subtractive Inhibition Assay for the Detection of E. coli O157:H7 Using Surface Plasmon Resonance |
| title_full | Subtractive Inhibition Assay for the Detection of E. coli O157:H7 Using Surface Plasmon Resonance |
| title_fullStr | Subtractive Inhibition Assay for the Detection of E. coli O157:H7 Using Surface Plasmon Resonance |
| title_full_unstemmed | Subtractive Inhibition Assay for the Detection of E. coli O157:H7 Using Surface Plasmon Resonance |
| title_short | Subtractive Inhibition Assay for the Detection of E. coli O157:H7 Using Surface Plasmon Resonance |
| title_sort | subtractive inhibition assay for the detection of e coli o157 h7 using surface plasmon resonance |
| topic | subtractive inhibition assay SPR E. coli O157:H7 |
| url | http://www.mdpi.com/1424-8220/11/3/2728/ |
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