Novel paternity testing by distinguishing parental alleles at a VNTR locus in the differentially methylated region upstream of the human H19 gene.
Conventional PCR-based genotyping is useful for forensic testing but cannot be used to determine parental origins of alleles in DNA specimens. Here we describe a novel method of combined conventional genotyping and PIA typing (parentally imprinted allele typing) at a minisatellite region upstream from the H19 locus. The PIA typing uses two sets of primers and DNA digested with methylation-sensitive Hha I enzyme. The first amplification produces only the methylated fragment of paternal H19 allele, and the second detects polymorphism in the minisatellite. Hence, this distinguishes paternal and maternal alleles by difference in the DNA methylation. Furthermore, the polymorphism in this polymorphic locus was examined using 199 unrelated Japanese and 171 unrelated Germans, their polymorphism information content being 0.671 and 0.705, respectively. Feasibility of this typing is demonstrated for six families, and the usefulness is shown by application to paternity testing.
Naito
Emiko
E
Division of Legal Medicine, Niigata University Graduate School of Medical and Dental Science, 1-757 Asahimachidori, Niigata 951-8510, Japan.
Dewa
Koji
K
Fukuda
Masaaki
M
Sumi
Hirokazu
H
Wakabayashi
Yui-ichi
Y
Umetsu
Kazuo
K
Yuasa
Isao
I
Yamanouchi
Haruo
H
D000483:Alleles; D016172:DNA Fingerprinting / Q000379:methods*; D019175:DNA Methylation; D017931:DNA Primers; D004586:Electrophoresis; D005006:Ethnicity / Q000235:genetics; D005260:Female; D005838:Genotype; D006801:Humans; D008297:Male; D018598:Minisatellite Repeats; D010334:Paternity; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic; D062085:RNA, Long Noncoding; D022661:RNA, Untranslated / Q000235:genetics*
D017931:DNA Primers; C094335:H19 long non-coding RNA; D062085:RNA, Long Noncoding; D022661:RNA, Untranslated