American Journal of BioScience

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Direct Determination of ABO Blood Group Genotypes from Whole Blood Using PCR-Amplification of Specific Alleles Method

Received: 14 February 2014    Accepted:     Published: 30 March 2014
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Abstract

ABO antigens are known to be oligosaccharide antigens, and are widely expressed on the membranes of red blood cells and tissue cells. Therefore, the determining ABO blood group antigens is important in both transfusion and organ transplantation, and is one of the markers used for personal identification in forensics. The ABO blood group is currently determined by the presence of A and B antigens on red blood cells using serological tests in clinical laboratories. The gene sequences of the ABO blood group in chromosome 9q34.1-q34.2 have also been determined. Accordingly, it has become possible to genetically analyze the ABO blood group using molecular biological techniques. We recently developed an ABO genotyping method based on PCR amplification of specific alleles (PASA) using DNA extracted from blood and saliva. However, the extraction and purification of DNA is necessary prior to PCR because blood and saliva contain various substances that inhibit PCR. Furthermore, the PCR amplification of specific alleles (PASA) method requires specificity and stability for allele-specific amplification. Therefore, it is very difficult to use whole blood directly. Here, we described the development and use of ABO genotyping from whole blood using a commercially available reagent kit, which can effectively neutralize inhibitory substances present in the blood. In this study, for all genes of the six major ABO genotypes and cisA2B3 genotype of the AB variant, only specific bands were clearly amplified, whereas non-specific bands were not amplified at all. In addition, this method was able to determine ABO genotyping using 5-fold diluted fresh whole blood, or 5-fold diluted whole blood that was freeze-stored in 100 µL aliquots at -20 oC by subdivided for a maximum of 30 days. This analysis method to determine ABO blood group genotyping is simple and useful, and is expected to be used widely throughout research and clinical laboratories and forensic fields.

DOI 10.11648/j.ajbio.20140202.15
Published in American Journal of BioScience (Volume 2, Issue 2, March 2014)
Page(s) 49-55
Creative Commons

This is an Open Access article, distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution and reproduction in any medium or format, provided the original work is properly cited.

Copyright

Copyright © The Author(s), 2024. Published by Science Publishing Group

Keywords

ABO Blood Group, Direct PCR, DNA Typing, PASA: PCR Amplification of Specific Alleles, CisAB

References
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Author Information
  • Department of Cells and Immunity Analytics, Institute of Health Biosciences, The University of Tokushima Graduate School, Tokushima, Japan

  • Department of Pharmacy, Tokushima University Hospital, Tokushima, Japan

  • Clinical Laboratory, Mie Prefectural Shima Hospital, Mie, Japan

  • Department of Cells and Immunity Analytics, Institute of Health Biosciences, The University of Tokushima Graduate School, Tokushima, Japan; Subdivision of Biomedical Laboratory Sciences, Graduate School of Health Sciences, The University of Tokushima, Tokushima, Japan

  • Department of Pharmacy, Tokushima University Hospital, Tokushima, Japan

  • Department of Cells and Immunity Analytics, Institute of Health Biosciences, The University of Tokushima Graduate School, Tokushima, Japan

Cite This Article
  • APA Style

    Kensaku Aki, Kazuyoshi Kawazoe, Azusa Izumi, Tomoki Tada, Kazuo Minakuchi, et al. (2014). Direct Determination of ABO Blood Group Genotypes from Whole Blood Using PCR-Amplification of Specific Alleles Method. American Journal of BioScience, 2(2), 49-55. https://doi.org/10.11648/j.ajbio.20140202.15

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    ACS Style

    Kensaku Aki; Kazuyoshi Kawazoe; Azusa Izumi; Tomoki Tada; Kazuo Minakuchi, et al. Direct Determination of ABO Blood Group Genotypes from Whole Blood Using PCR-Amplification of Specific Alleles Method. Am. J. BioScience 2014, 2(2), 49-55. doi: 10.11648/j.ajbio.20140202.15

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    AMA Style

    Kensaku Aki, Kazuyoshi Kawazoe, Azusa Izumi, Tomoki Tada, Kazuo Minakuchi, et al. Direct Determination of ABO Blood Group Genotypes from Whole Blood Using PCR-Amplification of Specific Alleles Method. Am J BioScience. 2014;2(2):49-55. doi: 10.11648/j.ajbio.20140202.15

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  • @article{10.11648/j.ajbio.20140202.15,
      author = {Kensaku Aki and Kazuyoshi Kawazoe and Azusa Izumi and Tomoki Tada and Kazuo Minakuchi and Eiji Hosoi},
      title = {Direct Determination of ABO Blood Group Genotypes from Whole Blood Using PCR-Amplification of Specific Alleles Method},
      journal = {American Journal of BioScience},
      volume = {2},
      number = {2},
      pages = {49-55},
      doi = {10.11648/j.ajbio.20140202.15},
      url = {https://doi.org/10.11648/j.ajbio.20140202.15},
      eprint = {https://download.sciencepg.com/pdf/10.11648.j.ajbio.20140202.15},
      abstract = {ABO antigens are known to be oligosaccharide antigens, and are widely expressed on the membranes of red blood cells and tissue cells. Therefore, the determining ABO blood group antigens is important in both transfusion and organ transplantation, and is one of the markers used for personal identification in forensics. The ABO blood group is currently determined by the presence of A and B antigens on red blood cells using serological tests in clinical laboratories. The gene sequences of the ABO blood group in chromosome 9q34.1-q34.2 have also been determined. Accordingly, it has become possible to genetically analyze the ABO blood group using molecular biological techniques. We recently developed an ABO genotyping method based on PCR amplification of specific alleles (PASA) using DNA extracted from blood and saliva. However, the extraction and purification of DNA is necessary prior to PCR because blood and saliva contain various substances that inhibit PCR. Furthermore, the PCR amplification of specific alleles (PASA) method requires specificity and stability for allele-specific amplification. Therefore, it is very difficult to use whole blood directly. Here, we described the development and use of ABO genotyping from whole blood using a commercially available reagent kit, which can effectively neutralize inhibitory substances present in the blood. In this study, for all genes of the six major ABO genotypes and cisA2B3 genotype of the AB variant, only specific bands were clearly amplified, whereas non-specific bands were not amplified at all. In addition, this method was able to determine ABO genotyping using 5-fold diluted fresh whole blood, or 5-fold diluted whole blood that was freeze-stored in 100 µL aliquots at -20 oC by subdivided for a maximum of 30 days. This analysis method to determine ABO blood group genotyping is simple and useful, and is expected to be used widely throughout research and clinical laboratories and forensic fields.},
     year = {2014}
    }
    

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  • TY  - JOUR
    T1  - Direct Determination of ABO Blood Group Genotypes from Whole Blood Using PCR-Amplification of Specific Alleles Method
    AU  - Kensaku Aki
    AU  - Kazuyoshi Kawazoe
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    AU  - Eiji Hosoi
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    JF  - American Journal of BioScience
    JO  - American Journal of BioScience
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    PB  - Science Publishing Group
    SN  - 2330-0167
    UR  - https://doi.org/10.11648/j.ajbio.20140202.15
    AB  - ABO antigens are known to be oligosaccharide antigens, and are widely expressed on the membranes of red blood cells and tissue cells. Therefore, the determining ABO blood group antigens is important in both transfusion and organ transplantation, and is one of the markers used for personal identification in forensics. The ABO blood group is currently determined by the presence of A and B antigens on red blood cells using serological tests in clinical laboratories. The gene sequences of the ABO blood group in chromosome 9q34.1-q34.2 have also been determined. Accordingly, it has become possible to genetically analyze the ABO blood group using molecular biological techniques. We recently developed an ABO genotyping method based on PCR amplification of specific alleles (PASA) using DNA extracted from blood and saliva. However, the extraction and purification of DNA is necessary prior to PCR because blood and saliva contain various substances that inhibit PCR. Furthermore, the PCR amplification of specific alleles (PASA) method requires specificity and stability for allele-specific amplification. Therefore, it is very difficult to use whole blood directly. Here, we described the development and use of ABO genotyping from whole blood using a commercially available reagent kit, which can effectively neutralize inhibitory substances present in the blood. In this study, for all genes of the six major ABO genotypes and cisA2B3 genotype of the AB variant, only specific bands were clearly amplified, whereas non-specific bands were not amplified at all. In addition, this method was able to determine ABO genotyping using 5-fold diluted fresh whole blood, or 5-fold diluted whole blood that was freeze-stored in 100 µL aliquots at -20 oC by subdivided for a maximum of 30 days. This analysis method to determine ABO blood group genotyping is simple and useful, and is expected to be used widely throughout research and clinical laboratories and forensic fields.
    VL  - 2
    IS  - 2
    ER  - 

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