Blood Grouping Based on PCR Methods and Agarose Gel Electrophoresis

  • Ana Maria Sell
  • Jeane Eliete Laguila Visentainer
Part of the Methods in Molecular Biology book series (MIMB, volume 1310)


The study of erythrocyte antigens continues to be an intense field of research, particularly after the development of molecular testing methods. More than 300 specificities have been described by the International Society for Blood Transfusion as belonging to 33 blood group systems. The polymerase chain reaction (PCR) is a central tool for red blood cells (RBC) genotyping. PCR and agarose gel electrophoresis are low cost, easy, and versatile in vitro methods for amplifying defined target DNA (RBC polymorphic region). Multiplex-PCR, AS-PCR (Specific Allele Polymerase Chain Reaction), and RFLP-PCR (Restriction Fragment Length Polymorphism-Polymerase Chain Reaction) techniques are usually to identify RBC polymorphisms. Furthermore, it is an easy methodology to implement. This chapter describes the PCR methodology and agarose gel electrophoresis to identify the polymorphisms of the Kell, Duffy, Kidd, and MNS blood group systems.

Key words

Blood group antigens Genotyping SNPs PCR Polymorphism 


  1. 1.
    Daniels G (2002) Human blood groups. Blackwell Science, OxfordCrossRefGoogle Scholar
  2. 2.
    Daniels G (2009) The molecular genetics of blood group polymorphism. Hum Genet 126:729–742PubMedGoogle Scholar
  3. 3.
    Colin Y, Chérif-Zahar B, Le Van Kim C et al (1991) Genetic basis of the RhD-positive and RhD-negative blood group polymorphism as determined by Southern analysis. Blood 78:2747–2752PubMedGoogle Scholar
  4. 4.
    Wagner T, Berer A, Lanzer G, Geissler K (2000) Kell is not restricted to the erythropoietic lineage but is also expressed on myeloid progenitor cells. Br J Haematol 110:409–411PubMedGoogle Scholar
  5. 5.
    Wagner T, Lanzer G, Geissler K (2002) Kell expression on myeloid progenitor cells. Leuk Lymphoma 43:479–485PubMedGoogle Scholar
  6. 6.
    Iwamoto S, Omi T, Kajii E, Ikemoto S (1995) Genomic organization of the glycoprotein D gene: Duffy blood group Fya/Fyb alloantigen system is associated with a polymorphism at the 44-amino acid residue. Blood 85:622–626PubMedGoogle Scholar
  7. 7.
    You G, Smith CP, Kanai Y et al (1993) Cloning and characterization of the vasopressin-regulated urea transporter. Nature 365:844–847CrossRefPubMedGoogle Scholar
  8. 8.
    Reid ME (2009) MNS blood group system: a review. Immunohematology 25:95–101PubMedGoogle Scholar
  9. 9.
    Strachan T, Read AP (1999) PCR, DNA sequencing and in vitro mutagenesis. Human molecular genetics. Wiley-Liss, New York, NYGoogle Scholar
  10. 10.
    Maaskant-Van Wijk PA, Faas BH, De Ruijter JA et al (1998) Genotyping of RHD by multiplex polymerase chain reaction analysis of six RHD-specific exons. Transfusion 38:1015–1021CrossRefPubMedGoogle Scholar
  11. 11.
    Singleton BK, Green CA, Avent ND et al (2000) The presence of on RHD pseudogene containing a 37 base pair duplication and a nonsense mutation in Africans with the Rh-negative blood group phenotype. Blood 95:12–18PubMedGoogle Scholar
  12. 12.
    Rios M, Cash K, Strupp A et al (1999) DNA from urine sediment or buccal cells can be used for blood group molecular genotyping. Immunohematology 15:61–65PubMedGoogle Scholar
  13. 13.
    Reid ME, Rios M, Yazdanbakhsh K (2000) Applications of molecular biology techniques to transfusion medicine. Semin Hematol 37:166–176PubMedGoogle Scholar
  14. 14.
    Castilho L, Rios M, Bianco C et al (2002) DNA-based typing for the management of multiply-transfused sickle cell disease patients. Transfusion 42:232–238CrossRefPubMedGoogle Scholar
  15. 15.
    Castilho L, Rios M, Pellegrino J Jr et al (2002) Blood group genotyping facilitates transfusion of beta-thalassemia patients. J Clin Lab Anal 16:216–220PubMedGoogle Scholar
  16. 16.
    Male DA, Phillips P, Turner DR (2002) Improved polymerase chain reaction-based glycophorin-A M/N blood group allelotyping. Vox Sang 83:170–171PubMedGoogle Scholar
  17. 17.
    Lavoiet J, Perreault J, St-Louis M (2008) Génotypage du systeme S et s par PCR en multiplex. Version 2. Département de Recherche et Développement, QuébecGoogle Scholar
  18. 18.
    Tournamille C, Colin Y, Cartron JP et al (1995) Disruption of a GATA motif in the Duffy gene promoter abolishes erythroid gene expression in Duffy-negative individuals. Nat Genet 10:224–228PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Ana Maria Sell
    • 1
  • Jeane Eliete Laguila Visentainer
    • 1
  1. 1.Laboratório de Imunogenética, Departamento de Ciências Básicas da SaúdeUniversidade Estadual de MaringáMaringáBrazil

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