A chemiluminescence derivatization method for detecting nucleic acids and DNA probes using a trimethoxyphenylglyoxal reagent that recognizes guanine

A chemiluminescence derivatization method using a novel reagent, 3',4',5'-trimethoxyphenylglyoxal (TMPG) was studied for the quantitative detection of nucleic acids and deoxyguanylic acid oligomers on a nylon membrane. In the method, TMPG rapidly converted guanine of a nucleic acid to a chemiluminescent derivative in an alkaline phosphate solution at a room temperature. A cooled, charge-coupled device camera was used to detect the chemiluminescence emitted at room temperature from the derivatives on the membrane after the TMPG reaction. The detection signal was increased by increasing the amount of deoxyguanylic acid in the analyte molecule. When a DNA probe having a 5'-(G)(15)TT(G)(15)TT deoxyoligomer at its 3' terminus was hybridized with a target complementary DNA fragment on the membrane, an increased chemiluminescence from the hybrid was detected by the proposed method. The lower limit of detection is approximately 1.0 pmol of the target DNA fragment bound to the probe in the hybridization assay. (C) 1999 Elsevier Science B.V. All rights reserved.