ABO Genotyping by Polymerase Chain Reaction

Abstract

ABO blood group system's genotyping by polymerase chain reaction in genomic DNA level is developed. The positions of nucleotide 258 and 700 of cDNA from A transferase were used to distinguish A, B, and O alleles by restriction enzyme digestion. To identify the 258th nucleotide, a 199- or 200-bp DNA fragment was amplified by PCR and digested with Kpn I. For the 700th nucleotide, a 128-bp PCR amplified fragment was designed and digested with Alu I. By examining the DNA fragment digested patterns, ABO genotypes were easily determined. Results obtained using this method on 20 ABO-known peripheral blood samples showed that this new technique could provide accurate ABO genotype. Biologic forensic samples, such as, blood stains, saliva stains, semen stains, hair, bone tissue, and semen contaminated with vaginal secretion were also successfully typed. This rapid, sensitive and reliable method should be applicable not only in forensic identification but also in medical examination.

Rapid ABO Genotyping Using Whole Blood without DNA Purification

“…There are several polymerase chain reaction (PCR) based methods used to identify the ABO genotypes, such as PCR followed by restriction digestion [3,4], denaturing gradient gel electrophoresis [5], single-strand conformation polymorphism [6], and allele-specific PCR [7,8]. Although PCR is technically simple, it is labor-intensive and time-consuming.…”

Donor DNA in a renal cell carcinoma metastasis from a bone marrow transplant recipient

“…9 Two primer pairs were designed to distinguish between A and O alleles (Table 1A). 10 The primer pairs were redundant and were used in different reaction tubes. Restriction fragments were generated by KpnI digestion.…”

O-gene detection by allele specific amplification in the ABO blood group system

“…Their primers were overlapped at numbers 237-258 bases for a sense primer and 257-276 bases for an anti-sense primer from O-gene cDNA and they used a complex reaction mixture and a radioisotope for detection of the PCR product. Yamamoto et al (1990b) and Lee and Chang (1992) reported PCR-RFLP method. This method was used most popularly at this time.…”

“…Determination of ABO genotype from peripheral blood leukocytes was first presented by Yamamoto et al (1990b). Several variations were developed by using PCR-RFLP method (Lee and Chang, 1992), allele specific PCR (AS-PCR) method (Ugozzoli and Wallace, 1992), and PCR-denaturing gradient gel electrophoresis (PCR-DGGE) method (Johnson and Hopkinson, 1992). Yamamoto's method was shown to be accurate in these three years.…”