ABO Genotyping of Forensic Specimens by PCR-RFLP Method Using Innovated Primers

Abstract

Innovated primers for ABO genotyping were designed. To identify the 261 G deletion in O allele, a 101- or 102-bp PCR fragment was amplified by 261-f and 261-r primers and digested with Kpn I. For the detection of the 703 point mutation (G→A) in B allele, a 131-bp PCR fragment was amplified by 703-f and 703-r primers and digested with Msp I. Results obtained with these primers for 6 ABO-known fresh peripheral blood samples showed that these new primers could provide an accurate ABO genotype. As for the amount of template DNA, ABO genotype could be performed with approximately 10 pg of template DNA. In forensic specimens (e.g., whole blood, skeletal muscle, hair and/or bone marrow) with different postmortem intervals between 7 hours and 7 years, it was possible to successfully determine ABO genotype.