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.
Published in | American Journal of BioScience (Volume 2, Issue 2) |
DOI | 10.11648/j.ajbio.20140202.15 |
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), 2014. Published by Science Publishing Group |
ABO Blood Group, Direct PCR, DNA Typing, PASA: PCR Amplification of Specific Alleles, CisAB
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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
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
@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://article.sciencepublishinggroup.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} }
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 AU - Azusa Izumi AU - Tomoki Tada AU - Kazuo Minakuchi AU - Eiji Hosoi Y1 - 2014/03/30 PY - 2014 N1 - https://doi.org/10.11648/j.ajbio.20140202.15 DO - 10.11648/j.ajbio.20140202.15 T2 - American Journal of BioScience JF - American Journal of BioScience JO - American Journal of BioScience SP - 49 EP - 55 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 -