Abstract
The genotyping of ABO blood groups was performed using the polymerase chain reaction (PCR) method. The 4 DNA fragments containing the nucleotide position 261, 526, 703 and 796 of cDNA from A-transferase were amplified by PCR, and the amplified DNA subjected to restriction fragment length polymorphism (RFLP) analysis. The different nucleotide at position 803 was clearly distinguished by electrophoresis of the PCR products amplified with allele-specific primers. By analyzing the electrophoresis patterns, ABO genotyping was conclusively accomplished. The frequencies of ABO genotypes found in Japanese blood donors with A and B phenotypes were as follows: in the phenotype A group, AA =19.8 % and AO = 80.2%; and in the phenotype B group, BB =12.8% and BO=87.2%.
Zusammenfassung
Die Bestimmung der ABO-Blutgruppen wurde mittels Polymerase-Kettenreaktion (PCR) durchgeführt. Die vier DNA-Fragmente, die Nukleotidpositionen 261, 526, 703 und 796 von der cDNA der A-Transferase enthielten, wurden mittels PCR amplifiziert. Die amplifizierte DNA wurde einer Restriktionsfragmentl ängen-Analyse (RFLP) unterzogen. Nach Amplifikation mit allelspezifischen Primern konnte der Nukleotidunterschied an Position 803 durch Elektrophorese der PCR-Produkte klar getrennt werden. Die Bestimmung der ABO-Genotypen war durch die Analyse der eletrophoretischen Muster eindeutig durchführbar. Folgende Frequenzen der ABO-Genotypen von japanischen Blutspendern wurden für die Phänotypen A und B gefunden: In der Phänotypgruppe A, AA =19,8% und AO=80,2%, Phänotypgruppe B, BB=12,8% und BO= 87,2%.
Similar content being viewed by others
References
Yamamoto F, Marken J, Tsuji T, White T, Clausen H, Hakomori S (1990) Cloning and characterization of DNA complementary to human UDP-Ga1NAc:Fucα1-2Galα1-3Ga1NAc transferase (histo-blood group A transferase) mRNA. J Biol Chem 265: 1146–1151
Yamamoto F, Clausen H, White T, Marken J, Hakomori S (1990) Molecular genetic basis of the histo-blood group ABO system. Nature 345: 229–233
Yamamoto F, Hakomori S (1990) Sugar-nucleotide donor specificity of histo-blood group A and B transferases is based on amino acid substitutions. J Biol Chem 165: 19257–19262
Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory, USA
Saiki RK, Scharf SJ, Faloona F, Mullis KB, Horn GT, Erlich HA, Arnheim N (1985) Enzymatic amplification of β-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia. Science 230: 1350–1354
Kaneshige T, Takagi K, Nakamura S, Hirasawa T, Sada M, Uchida K (1992) Genetic analysis using fingernail DNA. Nucleic Acids Res 20: 5489–5490
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Fukumori, Y., Ohnoki, S., Shibata, H. et al. Genotyping of ABO blood groups by PCR and RFLP analysis of 5 nucleotide positions. Int J Leg Med 107, 179–182 (1995). https://doi.org/10.1007/BF01428401
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF01428401