Korean J Blood Transfus.  2011 Aug;22(2):110-119.

Application of Real-Time PCR for ABO Genotyping for Large-scale Population Screening

Affiliations
  • 1Department of Laboratory Medicine, Chonnam National University Medical School, Gwangju, Korea. dcho@chonnam.ac.kr
  • 2Department of Preventive Medicince, Chonnam National University Medical School, Gwangju, Korea. ujingogo@paran.com
  • 3Blood Transfusion Research Institute, Korean Red Cross, Seoul, Korea.

Abstract

BACKGROUND
For large-scale population screening, the method of ABO genotyping needs to be simple, accurate and cost-effective. The real-time PCR method has been introduced and it is suitable for dealing with large numbers of specimens. In this study, we examined the ABO genotyping of 1,700 residents of Jeollanam-do for an epidemiologic study by applying the real-time PCR method.
METHODS
Genomic DNA was extracted from the peripheral blood samples of 1,700 residents of Jeollanam-do between July 2004 and January 2006 and these samples were stored at -70degrees C. The ABO genotype in all the samples was determined by four-color real-time PCR using displacing probes and three cases that had an atypical real time PCR pattern were confirmed by direct sequencing and PCR-based cloning of exons 6&7 of the ABO gene.
RESULTS
The genotyping results of 1,700 samples included O/O (25.6%), A/A (9.1%), A/O (29.1%), B/B (4.5%), B/O (19.8%) and A/B (11.9%), and the allele frequencies of O, A and B were 50.1%, 29.5% and 20.4%, respectively. The frequency of the O allele was lower in the residents of Jeollanam-do than that previously reported for the residents of Kangwon-do (P=0.014), while the frequency of the A allele was higher in the residents of Jeollanam-do than that previously reported for the residents of Kangwon-do (P=0.003). The three cases with atypical results were revealed to be B101/O24, Bvar(296C>T)/O01 and B101/Ovar(801G>T). It takes 6 days to perform ABO genotyping on 1,700 samples by a calculation per test.
CONCLUSION
ABO genotyping by real-time PCR using displacing probes can be useful for mass screening for ABO genotyping. In Korea, the frequency of the ABO allele was significantly different among different regions.

Keyword

ABO genotype; Real-time PCR; Direct DNA sequencing; Displacing probe; Fluorescence

MeSH Terms

Alleles
Clone Cells
Cloning, Organism
DNA
Epidemiologic Studies
Exons
Fluorescence
Gene Frequency
Genotype
Korea
Mass Screening
Real-Time Polymerase Chain Reaction
DNA
Full Text Links
  • KJBT
Actions
Cited
CITED
export Copy
Close
Share
  • Twitter
  • Facebook
Similar articles
Copyright © 2024 by Korean Association of Medical Journal Editors. All rights reserved.     E-mail: koreamed@kamje.or.kr