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Updated: Feb 8, 2026
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Polymerase Chain Reaction: Basic Protocol Plus Troubleshooting and Optimization Strategies
Published on: May 22, 2012
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Optimal Conditions for the Asymmetric Polymerase Chain Reaction for Detecting Food Pathogenic Bacteria Using a
Haruka Nagai1, Kanji Tomioka1, Shiro Okumura2
1Department of Biochemistry and Applied Chemistry, National Institute of Technology, Kurume College, 1-1-1 Komorino, Kurume, Fukuoka, 830-8555, Japan.
Applied Biochemistry and Biotechnology
|June 27, 2018
Summary
A new system uses asymmetric PCR and a portable SPR sensor for rapid foodborne pathogen detection. This method accurately quanties DNA amplicons, crucial for reliable point-of-care testing in the food industry.
Area of Science:
- Food safety
- Analytical chemistry
- Biotechnology
Background:
- Foodborne bacterial infections pose significant public health risks.
- Rapid and accurate detection methods are essential for the food industry.
- Existing detection systems may lack portability or sensitivity for field applications.
Purpose of the Study:
- To develop a quick and simple system for detecting foodborne pathogens.
- To establish a novel method for quantifying DNA amplicons using HPLC.
- To validate the correlation between DNA quantification and SPR sensor response.
Main Methods:
- Asymmetric PCR amplification of Staphylococcus aureus DNA.
- Quantification of forward (F) and reverse (R) DNA chains using High-Performance Liquid Chromatography (HPLC).
- Correction method for R-chain quantification considering F-chain concentration.
- Validation using fluorescence imaging and Surface Plasmon Resonance (SPR) sensor.
Main Results:
- A novel HPLC method accurately quantified F- and R-DNA chains separately.
- Calculated single-stranded DNA amplicon concentration showed good agreement with fluorescence imaging.
- SPR sensor response correlated well with the quantified single-strand F-chain concentration.
- The developed system demonstrated suitability for point-of-care detection.
Conclusions:
- The combined asymmetric PCR and portable SPR sensor system enables rapid detection of foodborne bacteria.
- Accurate DNA amplicon quantification via HPLC is critical for system sensitivity.
- This technology offers a promising solution for food safety monitoring in the field.
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