Korean J Lab Med.  2009 Jun;29(3):231-237. 10.3343/kjlm.2009.29.3.231.

Rapid ABO Genotyping Using Whole Blood without DNA Purification

• Lee SH
• Park G
• Yang YG
• Lee SG
• Kim SW

• KMID: 1096975
• DOI: http://doi.org/10.3343/kjlm.2009.29.3.231

Abstract

BACKGROUND: ABO genotyping is commonly used in cases of an ABO discrepancy between cell typing and serum typing, as well as in forensic practice for personal identification and paternity testing. We evaluated ABO genotyping via multiplex allele-specific PCR (ASPCR) amplification using whole blood samples without DNA purification.
METHODS
A four-reaction multiplex ASPCR genotyping assay was designed to detect specific nucleotide sequence differences between the six ABO alleles A101, A102, B101, O01, O02, and cis-AB01. The ABO genotypes of 127 randomly chosen samples were determined using the new multiplex ASPCR method.
RESULTS
The genotypes of the 127 samples were found to be A101/A102 (n=1), A102/A102 (n=9), A101/O01 (n=3), A102/O01 (n=12), A102/O02 (n=14), B101/B101 (n=5), B101/O01 (n=18), B101/O02 (n=15), O01/O01 (n=14), O02/O02 (n=8), O01/O02 (n=14) and A102/B101 (n=14), from which phenotypes were calculated to be A (n=39), B (n=38), O (n=36) and AB (n=14). The multiplex ASPCR assay results were compared with the serologically determined blood group phenotypes and genotypes determined by DNA sequencing, and there were no discrepancies.
CONCLUSIONS
This convenient multiplex ASPCR assay, performed using whole blood samples, provides a supplement to routine serological ABO typing and might also be useful in other genotyping applications.

Keyword

ABO genotyping; Whole blood; Allele-specific polymerase chain reaction

MeSH Terms

ABO Blood-Group System/*genetics
*Alleles
DNA/blood
Genotype
Humans
Polymerase Chain Reaction/*methods

Figure

• 

  Fig. 1. Primer positions for ABO genotyping. The key polymorphisms are indicated at positions, 261, 297, 467, and 803. Alleles O01 and O02 have a G deletion at nucleotide position 261 (the next nucleotide is shown in parentheses). Nucleotides at these positions were detected in PCR reactions using the appropriate primers as shown with arrows (given in Table 1).

• 

  Fig. 2. Electropherogram of selected ABO genotypes. The gel photograph showing the size difference of the amplified products was obtained from the samples with 13 independent genotypes. The internal control produces a distinct 500 bp band in all four reactions. Lane M, size marker (100-500 bp ladder); lane 1, PCR reaction 1; lane 2, PCR reaction 2; lane 3, PCR reaction 3; lane 4, PCR reaction 4.

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