Applicability of the parentally imprinted allele (PIA) typing of a VNTR upstream the H19 gene to forensic samples of different tissues.
The parentally imprinted allele (PIA) typing that we have recently developed determines parental alleles at a VNTR locus in the differentially methylated region upstream of the human H19 gene. The usefulness of this typing was demonstrated by its application to blood samples in paternity cases. However, its applicability to other tissue DNA remains to be tested. DNA samples from fifteen different postmortem tissues such as cerebrum, skeletal muscle and skin were examined, all of which were obtained from three autopsy cases 2-11h after death. DNA was digested with a methylation-sensitive HhaI enzyme and diluted solutions of the digests were subjected to the first PCR amplification, providing amplification of only the paternal H19 methylated allele. Subsequent VNTR typing was carried out for the amplified products to determine which allele was of paternal origin. No tissue-dependent difference was observed and all the samples examined, though degraded, were successfully used for determining the paternal allele. These results substantiate the usefulness of PIA typing in forensic examinations. Its application to two identity cases, a burned male body and a male body with adipocere formation, was also shown.
Sumi
Hirokazu
H
Division of Legal Medicine, Institute of Medicine and Dentistry, Niigata University Academic Assembly, 1-757 Asahimachidori, Niigata 951-8510, Japan.
Naito
Emiko
E
Dewa
Koji
K
Fukuda
Masaaki
M
Xu
Hong-De
HD
Yamanouchi
Haruo
H
D000483:Alleles; D004247:DNA / Q000032:analysis; D016172:DNA Fingerprinting / Q000379:methods*; D019175:DNA Methylation; D005260:Female; D005838:Genotype; D006801:Humans; D008297:Male; D010334:Paternity; D016133:Polymerase Chain Reaction; D062085:RNA, Long Noncoding; D022661:RNA, Untranslated / Q000235:genetics*
C094335:H19 long non-coding RNA; D062085:RNA, Long Noncoding; D022661:RNA, Untranslated; D004247:DNA