ネステッドPCR法を用いた食品中の特定原材料(小麦)の検出

DOI 日本農学文献記事索引 Web Site 被引用文献6件 参考文献5件

書誌事項

タイトル別名
  • Detection of Wheat as an Allergenic Substance in Food by a Nested PCR Method
  • ネステッド PCRホウ オ モチイタ ショクヒンチュウ ノ トクテイ ゲンザイリョウ コムギ ノ ケンシュツ

この論文をさがす

抄録

A nested PCR method was developed for the detection of DNAs extracted from allergenic substances (here, wheat) in food. Because of DNA fragmentation, detection of wheat-specific DNA extracted from food, such as retort pouch food, is very difficult. Therefore, to improve the sensitivity of detection, a nested PCR primer pair (Wtr01NE2-5' and Wtr10NE5-3': amplicon size 97 bp) was newly designed within the region of the PCR products amplified by the official Japanese primer pair (Wtr01-5' and Wtr10-3'; amplicon size 141 bp) for wheat. Genomic DNAs of seven kinds of commercial processed foods containing wheat, wheat flour and three kinds of wheat flours pressure-heated at 100, 121 and 131°C were extracted with a commercial ion-exchange type kit by modifying the Japanese official method. The nested PCR method involved two PCR procedures. First, PCR was performed by varying both the PCR reagents and cycling conditions of the Japanese official method. Second, PCR was performed using the first PCR products diluted 200-fold with TE buffer. The Japanese official method enabled detection of only four of the seven kinds of foods and three of the four kinds of flours (one sample was just a trace), while the nested PCR method detected all seven foods and all four flours. Investigation of the detectability of the four kinds of wheat flours depending on the size of the amplified fragment using five primer pairs showed that its size must be kept to less than approximately 100 bp. The nested PCR method significantly improved the sensitivity of detection of wheat-specific DNA.

収録刊行物

  • 食品衛生学雑誌

    食品衛生学雑誌 49 (1), 23-30, 2008

    公益社団法人 日本食品衛生学会

被引用文献 (6)*注記

もっと見る

参考文献 (5)*注記

もっと見る

キーワード

詳細情報 詳細情報について

問題の指摘

ページトップへ