論文

査読有り
2018年6月26日

Optimal Conditions for the Asymmetric Polymerase Chain Reaction for Detecting Food Pathogenic Bacteria Using a Personal SPR Sensor

Applied Biochemistry and Biotechnology
  • Haruka Nagai
  • ,
  • Kanji Tomioka
  • ,
  • Shiro Okumura

開始ページ
1
終了ページ
15
記述言語
英語
掲載種別
研究論文(学術雑誌)
DOI
10.1007/s12010-018-2819-y
出版者・発行元
Humana Press Inc.

We have been developing quick and simple system for detecting food-poisoning bacteria using a combination of an asymmetric PCR and a portable surface plasmon resonance (SPR) sensor. The system would be suitable for point-of-care detection of food-poisoning bacteria in the field of food industry. In this study, we established a novel method for quantifying the amplified forward (F) and reverse (R) chains of Staphylococcus aureus separately by high-performance liquid chromatography (HPLC). The concentration of single-stranded DNA amplicon excessively amplified, which is crucial for the system, could be calculated as the difference between those of the F- and R-chains. For the R-chain, a correction based on the F-chain concentration in the sample was used to obtain a more accurate value, because the determination of the R-chain concentration was affected by that of the coexisting F-chain. The concentration values were also determined by fluorescence imaging for electrophoresis gels of amplicons with FITC- or Cy5-conjugated primers, and they were in good agreement with the values by the HPLC. The measured concentration of the single-strand F-chain correlated well with the value of the SPR response against the probe that was a complementary sequence of the F-chain, immobilized on the sensor chip of the SPR sensor.

リンク情報
DOI
https://doi.org/10.1007/s12010-018-2819-y
PubMed
https://www.ncbi.nlm.nih.gov/pubmed/29943274

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