Yang B, Viscidi R, Yolken R
Department of Pediatrics, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205.
Anal Biochem. 1993 Sep;213(2):422-5. doi: 10.1006/abio.1993.1441.
A novel approach for evaluation of polymerase chain reaction product was described. A biotin-labeled DNA (or RNA) probe is hybridized to the digoxigenin-labeled target PCR product. The hybrids are retained on an anti-biotin antibody-coated plate. An anti-digoxigenin Fab conjugated with alkaline phosphatase is used for generating either a fluorescence or a chromagenic substance. Both fluorogenic and chromagenic systems can be used for quantitative measurement of the PCR product/probe hybrids. The method is specific, fast, nonisotopic, sensitive, quantitative, and practical.