Bioluminescent PCR-RFLP Enzyme-Linked Immunosorbent Assay for Analysis of Vitamin D Receptor Gene Polymorphism.

DOI PDF Web Site Web Site Web Site 被引用文献5件 参考文献36件

この論文をさがす

抄録

We developed a sensitive and rapid PCR-RFLP ELISA using acetate kinase (AK) and firefly luciferase as a detection system. AK used as a label enzyme could sensitively be detected by bioluminescent assay using the firefly luciferase reac-tion. The detection limit was 10-20 mol/assay and the luminescence was stable for 48 h. FITC-labeled sense primer and biotin labeled anti sense primer were used for PCR amplification of the vitamin D receptor gene. After PCR, the products were digested with Taq I or Apa I enzyme. The reaction products were diluted with assay buffer and transferred to a plate coated with anti FITC IgG. After incubation for 2 h at 37°C, the plate was washed and reacted with avidin/biotinylated AK, the AK activity was detected by bioluminescence assay using the firefly luciferin/luciferase system. DNA polymorphism types (AA, Aa, aa, TT, Tt, tt) of the vitamin D receptor gene (VDR) could be clearly determined by measuring the bioluminescent intensity or by using photon imaging with a CCD camera.

収録刊行物

  • Analytical Sciences

    Analytical Sciences 15 (10), 943-949, 1999

    社団法人 日本分析化学会

被引用文献 (5)*注記

もっと見る

参考文献 (36)*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