We have developed a new non-isotopic DNA detection method. This method has 3 characteristics. First, a plant growth hormone, gibberellin, and its derivatives (GAs) are used as substances for labeling the DNA probe. Second, a novel fluorogenic substrate for alkaline phosphatase HNPP (2-hydroxy-3-naphthoic acid-2'-phenylanilide phosphate) has been synthesized, and GAs as a marker is detected by binding an alkaline phosphatase and anti-GA antibody conjugate. Third, a fluorescent image is recorded and the intensity is measured with an epi-fluorescence original detector, Epi-Light UV FA 1100, using a charged coupled device (CCD) camera that is designed to detect the signal from HNPP highly efficiently. The combination of the new fluorogenic substrate and the detector makes it possible to detect DNA on such a reaction support as a nylon membrane or in the in-situ reaction at a high resolutional level and high sensitivity. This new total detection method enables the development of a rapid, stable and highly sensitive system and will contribute to molecular biology and clinical diagnostics.
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