The Use of a Chemiluminescent Detection System for Paternity and Forensic Testing. Verification of the Reliability of the Oligonucleotide-Probes Used for Genetic Analysis

Summary

Several VNTR loci, that had been characterized by the use of P³²-labeled recombinant DNA fragments or oligonucleotides to the consensus sequence, were analyzed using oligonucleotide-Alkaline Phosphatase conjugates and a chemiluminescent detection method. The loci tested were D2S44, D4S163, D12S11, D14S13, D17S79, D18S27, DXYS14 and DNF24. Individuals, already typed using P³²-labeled probes, were used in this study. Pst I or Hae III-digested DNA from approximately 200 to 400 individuals, covering the full size range of alleles, were examined for each locus. The results obtained for these loci show that, with the exception of D2S44, all the alleles were recognized with the AP-oligos. D2S44 contained a subset of alleles that could only be detected by hybridization to recombinant DNA fragments. In forensic applications, results obtained using DNA recovered from evidentiary material show that most probes detect 10 ng of genomic DNA after an overnight exposure. This indicates that the sensitivity of this detection system is equal or better to that obtained with P³²-labeled probes.