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Clinical Chemistry 42: 14-18, 1996;
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Clinical Chemistry, Vol 42, 14-18, Copyright © 1996 by American Association for Clinical Chemistry

Chemiluminescent hybridization-ligation assays for delta F508 and delta I507 cystic fibrosis mutations

RA Martinelli, JC Arruda and P Dwivedi
Ciba-Corning Diagnostics Corp., E. Walpole, MA 02032, USA.

Chemiluminescent hybridization-ligation assays were devised to detect the delta F508 and delta I507 cystic fibrosis mutations in samples of human DNA that had been amplified by PCR. Two synthetic DNA oligomers were used in each assay. One of the oligomers was labeled with an acridinium ester and the other was immobilized on paramagnetic particles. The oligomers were hybridized to the samples and the target sequences discriminated by ligation with T4 or a thermostable DNA ligase. The performance of the assay was evaluated in a blind study of 30 samples. There was complete correspondence between the assignments based on the chemiluminescent assay and those made previously by gel electrophoresis, with one exception. The assignment of this discrepant sample by the chemiluminescent assay as a delta I507/normal heterozygote rather than a delta F508/normal heterozygote was confirmed by sequencing. The chemiluminescent hybridization-ligation assay provides a rapid and convenient means of discriminating DNA sequences differing by a single nucleotide.




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Copyright © 1996 by the American Association for Clinical Chemistry.