Problems of Forensic Sciences 2001 Vol. 48 (XLVIII) 53-64
GENOTYPING OF AB0 BLOOD GROUP ALLELES IN BLOODSTAINS BY THE PCR-RFLP TECHNIQUE
Marzanna CIESIELKA, Piotr KOZIOŁ, Roman MĄDRO
Chair and Department of Forensic Medicine, Medical Academy, Lublin
Streszczenie
Genotyping of the AB0 blood group system was performed using the method described by Lee and Chang, which enables differentiation of six AB0 genotypes: A0, AA, B0, BB, 00, AB. Amplification of DNA fragments 199–200 bp and 128 bp long (performed in one or two PCR reactions) was followed by restriction endonuclease Alu I and Kpn I digestion. Products of restriction digestion were subjected to separation in polyacrylamide gels and silver stained. Parameters of amplification, using two pairs of primers simultaneously and for each pair of primers separately, are described. The genotyping method was verified by studying 20 experimental stains (prepared from fresh blood) and 32 stains of blood collected during autopsies (prepared 2, 5, 10 and 15 years ago) of known blood group, which was determined earlier using serological methods. For all analysed experimental bloodstains, positive results of amplification with an input of 3 ng of DNA template and a correct result for AB0 blood grouping were obtained.
Słowa kluczowe
AB0; Genotyping; PCR-RFLP; Bloodstains.