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Label-free and high-sensitive detection of Salmonella using a surface plasmon resonance DNA-based biosensor

Paper ID Volume ID Publish Year Pages File Format Full-Text
23852 43478 2012 6 PDF Available
Title
Label-free and high-sensitive detection of Salmonella using a surface plasmon resonance DNA-based biosensor
Abstract

A method based on surface plasmon resonance (SPR) DNA biosensor has been developed for label-free and high-sensitive detection of Salmonella. A biotinylated single-stranded oligonucleotide probe was designed to target a specific sequence in the invA gene of Salmonella and then immobilized onto a streptavidin coated dextran sensor surface. The invA gene was isolated from bacterial cultures and amplified using a modified semi-nested asymmetric polymerase chain reaction (PCR) technique. In order to investigate the hybridization detection, experiments with different concentration of synthetic target DNA sequences have been performed. The calibration curve of synthetic target DNA had good linearity from 5 nM to 1000 nM with a detection limit of 0.5 nM. The proposed method was applied successfully to the detection of single-stranded invA amplicons from three serovars of Salmonella, i.e., Typhimurium, Enterica and Derby, and the responses to PCR products were related to different S. typhimurium concentrations in the range from 102 to 1010 CFU mL−1. While with this system to detect E. coli and S. aureus, no significant signal was observed, demonstrating good selectivity of the method. In addition, the hybridization can be completed within 15 min, and the excellent sensor surface regeneration allows at least 300 assay cycles without obvious loss of performance.

► A label-free and high-sensitive method based on surface plasmon resonance DNA biosensor has been proposed for detection of Salmonella. ► A modified asymmetric PCR has been introduced to produce predominantly ssDNA target. ► 102 CFU mL−1 of Salmonella cells form culture samples can be detected within 4.5 h. ► No significant signal was observed to detect E. coli and S. aureus.

Keywords
invA gene; Surface plasmon resonance; Biosensor; Polymerase chain reaction; Salmonella
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Label-free and high-sensitive detection of Salmonella using a surface plasmon resonance DNA-based biosensor
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 160, Issues 3–4, 31 August 2012, Pages 123–128
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us