Accurate Rh phenotype determination by reticulocyte mRNA typing shortly after multiple transfusions

This study describes a method for correctly typing patients’ own Rh-antigens after multiple transfusions. Alloimmunization is a common phenomenon after transfusion, with an estimated incidence of 0.5% increasing to 20–60% in chronically transfused patients. In recently transfused patients, serological typing can be hampered by mixed field agglutination. We established RT-PCR methods for RHD, RHC/c and RHE/e typing using mRNA from reticulocytes. Molecular typing was performed soon after 51 separate mismatched transfusion events involving 30 patients. Accurate identification of the transfused patients’ phenotype was confirmed in all cases. Reticulocyte maturation studies revealed that temperature is a crucial parameter for transition into mature red blood cells.

[1]  C. Drexler,et al.  Blood group chimerism , 2006, Current opinion in hematology.

[2]  L. Fast Microchimerism: a lasting legacy of transfusion? , 2006, Transfusion.

[3]  F. Claas Chimerism as a tool to induce clinical transplantation tolerance. , 2004, Current opinion in immunology.

[4]  Tzong-Hae Lee,et al.  Clinical investigation of posttransfusion Kidd blood group typing using a rapid normalized quantitative polymerase chain reaction , 2004, Transfusion.

[5]  I. Randen,et al.  Rapid and reliable genotyping of human platelet antigen (HPA)‐1, ‐2, ‐3, ‐4, and ‐5 a/b and Gov a/b by melting curve analysis , 2003, Transfusion.

[6]  F. Costa,et al.  DNA‐based typing of blood groups for the management of multiply‐transfused sickle cell disease patients , 2002, Transfusion.

[7]  C. Gassner,et al.  Differentiation of autologous ABO, RHD, RHCE, KEL, JK, and FY blood group genotypes by analysis of peripheral blood samples of patients who have recently received multiple transfusions , 2000, Transfusion.

[8]  M. E. Reid,et al.  DNA from blood samples can be used to genotype patients who have recently received a transfusion , 2000, Transfusion.

[9]  M. Lakomek,et al.  Application of RHD and RHCE genotyping for correct blood group determination in chronically transfused patients , 1999, Transfusion.

[10]  Gassner,et al.  Rh phenotype prediction by DNA typing and its application to practice , 1998, Transfusion medicine.

[11]  W. Flegel,et al.  RHD/CE typing by polymerase chain reaction using sequence‐specific primers , 1997, Transfusion.

[12]  R. H. Moore,et al.  In vitro and in vivo persistence of reticulocytes from donor red cells , 1996, Transfusion.

[13]  B. Carritt,et al.  DNA‐Based Rhesus Typing: Simultaneous Determination of RHC and RHD Status Using the Polymerase Chain Reaction , 1996, Vox sanguinis.

[14]  S. Hmida,et al.  Red cell alloantibodies in patients with haemoglobinopathies. , 1994, Nouvelle revue francaise d'hematologie.

[15]  J. Peristeri,et al.  Red Cell Alloantibodies in Patients with Thalassemia , 1990, Vox sanguinis.

[16]  P. Seligman,et al.  Automated analysis of reticulocytes using fluorescent staining with both acridine orange and an immunofluorescence technique , 1983, American journal of hematology.