A new method for human ABO genotyping using a universal reporter primer system.
We developed a new method for forensic ABO genotyping based on a universal reporter primer (URP) system. This allows for the simultaneous detection of six single nucleotide polymorphism (SNP) sites in the ABO gene (nucleotide positions 261, 297, 526, 703, 796, and 803). This URP system provides obvious peaks, ranging from 82 to 151 bp in length. ABO genotypes were classified and successfully genotyped by our method, including minor alleles that may cause a discrepancy between the genetic data and serological phenotypes. Full profiles were identified using as little as 0.1 ng (0.05 ng ⁄ reaction) of standard K562 and 9947A DNA. Moreover, the success rate of genotyping from a URP system was much higher than that from a conventional primer extension method in degraded DNA. This method enables simple and rapid detection of multiple SNP sites on human ABO genes and is highly specific and sensitive when using limited and degraded DNA.
Asari
Masaru
M
Department of Legal Medicine, Asahikawa Medical College, Asahikawa, 078-8510 Japan. asari@asahikawa-med.ac.jp
Omura
Tomohiro
T
Maseda
Chikatoshi
C
Matsubara
Kazuo
K
Shiono
Hiroshi
H
Shimizu
Keiko
K
D000017:ABO Blood-Group System / Q000235:genetics*; D053836:DNA Degradation, Necrotic; D017931:DNA Primers; D004586:Electrophoresis; D053803:Forensic Genetics / Q000379:methods*; D005838:Genotype; D006801:Humans; D007564:Japan; D016133:Polymerase Chain Reaction; D020641:Polymorphism, Single Nucleotide; D017422:Sequence Analysis, DNA
D000017:ABO Blood-Group System; D017931:DNA Primers