Quantitative detection of Staphylococcal Enterotoxin B by resonant acoustic profiling

Natesan, Mohan, Cooper, Matthew A., Tran, Julie P., Rivera, Victor R. and Poli, Mark A. (2009) Quantitative detection of Staphylococcal Enterotoxin B by resonant acoustic profiling. Analytical Chemistry, 81 10: 3896-3902. doi:10.1021/ac900086t


Author Natesan, Mohan
Cooper, Matthew A.
Tran, Julie P.
Rivera, Victor R.
Poli, Mark A.
Title Quantitative detection of Staphylococcal Enterotoxin B by resonant acoustic profiling
Formatted title
Quantitative detection of Staphylococcal Enterotoxin B by resonant acoustic profiling
Journal name Analytical Chemistry   Check publisher's open access policy
ISSN 0003-2700
Publication date 2009-05-15
Year available 2009
Sub-type Article (original research)
DOI 10.1021/ac900086t
Volume 81
Issue 10
Start page 3896
End page 3902
Total pages 7
Editor Royce W Murray
Place of publication Washington , DC, U.S.A.
Publisher American Chemical Society
Collection year 2010
Language eng
Subject 03 Chemical Sciences
0304 Medicinal and Biomolecular Chemistry
C1
861503 Scientific Instruments
020301 Acoustics and Acoustical Devices; Waves
Abstract A rapid and sensitive detection of staphylococcal enterotoxin B (SEB) was developed using a novel acoustic sensing technique: Resonant Acoustic Profiling (RAP), which utilizes high-frequency piezoelectric quartz resonators for monitoring biomolecular interactions. An automated four-channel instrument consisting of acoustic sensors covalently conjugated with anti-SEB antibodies was used. As the samples flowed across control and active sensors simultaneously, binding was measured as a change in the resonant frequency. The lower limit of detection (LLOD) for the label free direct format was 25 ng/mL. Detection sensitivity was increased by adding mass sequentially to the captured SEB on the sensor in the form of sandwich antibodies and biotin-avidin-based gold nanoparticles. The LLOD for the mass enhanced formats were 5 and 0.5 ng/mL of SEB, respectively. The lowest sensitivity corresponds to 1.3 fM in a 75 μL sample. The total assay time including the enhancement steps was less than 10 min. SEB was detected in both neat urine and PBS buffer-spiked samples, with linear correlations between resonant frequency signals and SEB concentrations (R2 of 0.999 and 0.998, respectively). No significant cross-reactivity was observed with homologue toxins SEA, SED, and TSST, but some crossreactivity was observed with the closely related toxin SEC1 when we used a polyclonal antibody in the assay. SEC1 cross-reactivity was not observed when a SEBspecific monoclonal antibody was employed in the assay. Thus the specificity of the assay presented here was dependent on the quality of the antibodies used. In addition to detection, we evaluated RAP’s ability to measure the toxin in unknown samples rapidly by measuring the initial binding rate of the interaction, thereby further shortening the assay time to 6 min. c 2009 American Chemical Society
Keyword Shock-syndrome toxin
Piezoelectric crystal immunosensor
Surface-plasmon resonance
Sensitive detection
DNA Hybridization
Human-antibodies
Microbalance
Biosensor
Superantigens
Localization
Q-Index Code C1
Q-Index Status Confirmed Code

 
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Created: Tue, 17 Nov 2009, 12:17:40 EST