Development of a chemiluminescent reagent that recognizes guanine in nucleic acid, and its use for chemiluminescence detection of DNA

一种新的试剂，3',4',5'-三甲氧基苯乙二醛，可以特异性地将核酸中的鸟嘌呤碱基在中性或碱性磷酸盐溶液中转化为相应的化学发光衍生物。化学致发光衍生化技术为脱氧核鸟苷酸低聚物DNA探针尾端的灵敏成像检测提供了一种简便的方法。冷却电荷耦合器件摄像机可用于视觉检测衍生化探针发出的化学发光，该探针与尼龙膜上的目标DNA片段杂交。所提出的方法可以定量检测探针在3‘端共价连接5’- (G) _<15>TT (G) _<15>TT脱氧寡聚物，表明在膜上的检测限为0.1 pmol。探针分子中脱氧核鸟苷酸的含量越大，检测灵敏度越高。

A novel reagent, 3', 4', 5'-trimethoxyphenylglyoxal has been found to specifically convert guanine bases of a nucleic acid in a neutral or alkaline phosphate solution to the corresponding chemiluminescent derivatives. The chemiluminogenic derivatization technique offers a facile method of the sensitive imaging detection for a DNA probe tailing with deoxyriboguanylic acid oligomers. A cooled charge-coupled device camera is usable to detect visually the chemiluminescence emitted from the derivatized probe that was hybridized with a target DNA fragment on a Nylon membrane. The proposed method permits the quantitative detection of the probe covalently attached with a 5'- (G) _<15>TT (G) _<15>TT deoxyoligomer at 3' terminus, indicating a detection limit of 0.1 pmol on the membrane. The detection sensitivity can be increased with larger contents of deoxyriboguanylic acid in the probe molecule.