Determination of ABO genotypes by real-time PCR using allele-specific primers

Muro, Tomonori; Fujihara, Junko; Imamura, Shinji; Nakamura, Hiroaki; Kimura-Kataoka, Kaori; Toga, Tomoko; Iida, Reiko; Yasuda, Toshihiro; Takeshita, Haruo

doi:10.1016/j.legalmed.2011.10.002

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Volume 14, Issue 1 , Pages 47-50, January 2012

Determination of ABO genotypes by real-time PCR using allele-specific primers

Tomonori Muro
- Department of Legal Medicine, Faculty of Medicine, Shimane University, Izumo 693-8501, Japan
- Criminal Investigation Laboratory, Shimane Prefectural Police Headquarters, Matsue 690-8510, Japan
Junko Fujihara
- Department of Legal Medicine, Faculty of Medicine, Shimane University, Izumo 693-8501, Japan
Shinji Imamura
- Department of Legal Medicine, Faculty of Medicine, Shimane University, Izumo 693-8501, Japan
- Criminal Investigation Laboratory, Shimane Prefectural Police Headquarters, Matsue 690-8510, Japan
Hiroaki Nakamura
- Criminal Investigation Laboratory, Shimane Prefectural Police Headquarters, Matsue 690-8510, Japan
Kaori Kimura-Kataoka
- Department of Legal Medicine, Faculty of Medicine, Shimane University, Izumo 693-8501, Japan
Tomoko Toga
- Department of Legal Medicine, Faculty of Medicine, Shimane University, Izumo 693-8501, Japan
Reiko Iida
- Division of Life Sciences, Faculty of Medical Sciences, University of Fukui, Fukui 910-1193, Japan
Toshihiro Yasuda
- Division of Medical Genetics and Biochemistry, Faculty of Medical Sciences, University of Fukui, Fukui 910-1193, Japan
Haruo Takeshita
- Department of Legal Medicine, Faculty of Medicine, Shimane University, Izumo 693-8501, Japan
- Corresponding author. Tel.: +81 853 20 2156; fax: +81 853 20 2155.

Received 31 October 2010; received in revised form 4 October 2011; accepted 18 October 2011. published online 19 December 2011.

Abstract

ABO grouping of biological specimens is informative for identifying victims and narrowing down suspects. In Japan and elsewhere, ABO grouping as well as DNA profiling plays an essential role in crime investigations. In the present study, we developed a new method for ABO genotyping using allele-specific primers and real-time PCR. The method allows for the detection of three single nucleotide polymorphisms (SNPs) at nucleotide positions 261, 796, and 803 in the ABO gene and the determination of six major ABO genotypes. This method required less than 2h for accurate ABO genotyping using 2.0ng of DNA. This method could be applicable for rapid and simple screening of forensic samples.