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Quantitative measurement of nonisotopically labeled polymerase chain reaction product
1Department of Pediatrics, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205.
Analytical Biochemistry
|September 1, 1993
Abstract:
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.