Automated DNA diagnostics using an ELISA-based oligonucleotide ligation assay
DNA diagnostics, the detection of specific DNA sequences, will play an increasingly important role in medicine as the molecular basis of human disease is defined. Here, we demonstrate an automated, nonisotopic strategy for DNA diagnostics using amplification of target DNA segments by the polymerase chain reaction (PCR) and the discrimination of allelic sequence variants by a colorimetric oligonucleotide ligation assay (OLA). We have applied the automated PCR/OLA procedure to diagnosis of common genetic diseases, such as sickle cell anemia and cystic fibrosis (delta F508 mutation), and to genetic linkage mapping of gene segments in the human T-cell receptor beta-chain locus. The automated PCR/OLA strategy provides a rapid system for diagnosis of genetic, malignant, and infectious diseases as well as a powerful approach to genetic linkage mapping of chromosomes and forensic DNA typing.
Additional Information© 1990 National Academy of Sciences. Contributed by Leroy Hood, August 16, 1990. We thank Ms. Anna Marie Aquinaldo and Drs. Conrad Sevilla and Suzanna Horvath for their assistance in oligonucleotide synthesis; all our cheek scraping donors; and Drs. P. Charmley, C. Delahunty, T. Hunkapiller, B. Koop, M. Nishimura, L. Rowen, and D. Zaller for their careful review of the manuscript. This work was supported by the Whittier Foundation, National Science Foundation Grant DIR 8809710, and National Institutes of Health Grant HG 00084.
Published - PNAS-1990-Nickerson-8923-7.pdf