CHEMILUMINESCENT HYBRIDIZATION-LIGATION ASSAYS FOR DELTA-F508 AND DELTA-1507 CYSTIC-FIBROSIS MUTATIONS

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 DIVA oligomers were used in each assay, One of the oligomers was labeled with an acr idinium 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 th e chemiluminescent assay and those made previously by gel electrophore sis, with one exception, The assignment of this discrepant sample by t he chemiluminescent assay as a Delta I507/normal heterozygote rather t han a Delta F508/normal heterozygote was confirmed by sequencing. The chemiluminescent hybridization-ligation assay provides a rapid and con venient means of discriminating DNA sequences differing by a single nu cleotide.