title: A simple ABO genotyping by PCR using sequence-specific primers with mismatched nucleotides.
abstract: In forensics, the specific ABO blood group is often determined by analyzing the ABO gene. Among various methods used, PCR employing sequence-specific primers (PCR-SSP) is simpler than other methods for ABO typing. When performing the PCR-SSP, the pseudo-positive signals often lead to errors in ABO typing. We introduced mismatched nucleotides at the second and the third positions from the 3'-end of the primers for the PCR-SSP method and examined whether reliable typing could be achieved by suppressing pseudo-positive signals. Genomic DNA was extracted from nail clippings of 27 volunteers, and the ABO gene was examined with PCR-SSP employing primers with and without mismatched nucleotides. The ABO blood group of the nail clippings was also analyzed serologically, and these results were compared with those obtained using PCR-SSP. When mismatched primers were employed for amplification, the results of the ABO typing matched with those obtained by the serological method. When primers without mismatched nucleotides were used for PCR-SSP, pseudo-positive signals were observed. Thus our method may be used for achieving more reliable ABO typing.
authors.affiliation: Department of Legal Medicine, School of Medicine, Tokyo Women's Medical University, 8-1 Kawada-cho, Shinjuku-ku, Tokyo 162-8666, Japan. Electronic address: taki@research.twmu.ac.jp.
authors.forename: Takashi
authors.initials: T
authors.lastname: Taki
authors.affiliation: Department of Legal Medicine, School of Medicine, Tokyo Women's Medical University, 8-1 Kawada-cho, Shinjuku-ku, Tokyo 162-8666, Japan.
authors.forename: Kazuhiko
authors.initials: K
authors.lastname: Kibayashi
mesh_terms: D000017:ABO Blood-Group System / Q000032:analysis / Q000235:genetics*; D001788:Blood Grouping and Crossmatching / Q000379:methods*; D017931:DNA Primers; D060005:Genotyping Techniques / Q000379:methods*; D006801:Humans; D009711:Nucleotides; D016133:Polymerase Chain Reaction / Q000379:methods*
chemical_list: D000017:ABO Blood-Group System; D017931:DNA Primers; D009711:Nucleotides
keywords: ABO genotyping; Allele-specific PCR; Pseudo-positive signal