Red Blood Cell Antigen Genotyping

Last Literature Review: October 2018 Last Update:
  • Use to predict the expression of RBC antigen specificities, aid in the selection of antigen-negative RBCs for transfusion, assess the risk of alloimmune hemolytic disease, or resolve conflicting serologic antibody results.
  • Detects 24 variants associated with 35 RBC antigens and phenotypic variants, in addition to the hemoglobin S variant.
  • For fetal testing, order Red Blood Cell Antigen Genotyping, Fetal (3016639) using a fetal specimen.

See Related Tests below for antigen-specific genotyping and phenotyping tests.

Red blood cell (RBC) antigen testing is useful in determining allelic variants that predict RBC antigen phenotypes for patients with recent history of transfusion or with conflicting serologic antibody results due to partial, variant, or weak expression antigens. It is also useful as an aid in management of hemolytic disease of the fetus and newborn (HDFN); for additional information on HDFN testing, refer to the ARUP Consult Hemolytic Disease of the Fetus and Newborn topic.

Disease Overview

Prevalence and/or Incidence

Erythrocyte alloimmunization occurs in up to 58% of patients with sickle cell, up to 35% in other patients who are transfusion-dependent, and in approximately 0.8% of all individuals who are pregnant.

Symptoms

Transfusion reactions or HDFN can occur due to alloimmunization:

  • Intravascular hemolysis: hemoglobinuria, jaundice, shock
  • Extravascular hemolysis: fever and chills
  • HDFN: fetal hemolytic anemia, hepatosplenomegaly, jaundice, erythroblastosis, neurologic damage, hydrops fetalis

Clinical presentation is variable and dependent upon the specific antibody and recipient factors.

Pathophysiology

Extravascular Hemolysis

Removal of antibody-bound RBCs via splenic macrophages

Intravascular Hemolysis

  • Rare for non-ABO antibodies
  • Donor RBCs are destroyed by the recipient's antibodies while they are still inside blood vessels.
  • Hemoglobin released into plasma and excreted in urine; bilirubin accumulates in blood

HDN-Mediated Disease

  • Caused by antigen-antibody mediated red-cell hemolysis from previously transferred maternal antibodies
  • Examples of antibodies associated with HDFN:
    • Rh (Ant-D, -C, -c, -E, and -e)
    • Kell (Anti-K, -k, -Kpa, -Kpb, -Jsa and -Jsb)
    • Duffy (Anti-Fya and -Fyb)
    • MNS (Anti-M, -N, -S, -s, and -U)
    • Kidd (Anti-Jka and -Jkb)
  • Anti-D is the most common cause of HDFN, followed by anti-c, anti-K, and anti-E; anti-Jka and -Jkb are rare causes
  • Antibodies cross the placenta and cause immune destruction of RBCs in fetus and leads to hydrops fetalis.

Genetics

Refer to the Genes and Variants Tested table below.

Inheritance

Typically codominant for RBC antigens; autosomal recessive for HbS

Test Interpretation

Sensitivity/Specificity

  • Clinical sensitivity and specificity:
    • >99% for C, c, E, e, K, k, Jka, Jkb, Fya, Fyb, M, N, S, s
    • Unknown for Kpa, Kpb, Jsa, Jsb, Lua, Lub, Dia, Dib, Coa, Cob, Doa, Dob, Joa, Hy, LWa, LWb, Sc1, Sc2, U, V, VS, HbS
  • Analytic sensitivity and specificity:
    • >99% for C, c, E, e, K, k, Jka, Jkb, Fya, Fyb, M, N, S, s
    • Unknown for Kpa, Kpb, Jsa, Jsb, Lua, Lub, Dia, Dib, Coa, Cob, Doa, Dob, Joa, Hy, LWa, LWb, Sc1, Sc2, U, V, VS, HbS

Results

  • Predicted phenotype for each RBC antigen tested will be provided
    • If identified, weak alleles will be reported.
  • Possible variant
    • Reported for allele combinations that have not widely been reported in the literature
  • Indeterminate
    • Abnormal signal intensities may result in the inability to predict genotype resulting in indeterminate results for all tested antigens and HbS.

Limitations

  • Only the variants listed will be interrogated.
  • Does not test for Rh D, a major cause of HDFN
  • Patients who have had allogeneic hematopoietic stem cell transplants may have inconclusive results.
  • Diagnostic/phenotype prediction errors can occur due to:
    • Rare variants affecting primer/probe binding
    • Molecular events that affect the blood-group antigen expression and phenotypes that are not detected by this assay (i.e., certain null phenotypes or other phenotypes with altered expression)
  • This assay is not designed to diagnose sickle cell disease.
Genes and Variants Tested
Blood GroupAlleleAntigen (ISBT #)ISBT GenotypeVariants TestedVariants Used to Predict AllelePhenotype Frequency , 
RhCRH2RHCE*2

c.307C>T; p.Pro103Ser

109bp insertion

c.307T; p.Ser103

109bp insertion

W: 68%

AA: 27%

A: 93%

cRH4RHCE*4c.307C; p.Pro103

W: 80%

AA: 98%

A: 47%

ERH3RHCE*3c.676G>C; p.Ala226Proc.676C; p.Pro226

W: 29%

AA: 22%

A: 39%

eRH5RHCE*5c.676G; p.Ala226

W: 98%

AA: 98%

A: 96%

VRH10

RHCE*01.20.01

RHCE*01.20.02

RHCE*01.20.04

RHCE*01.20.05

c.733C>G; p.Leu245Val

c.1006G>T; p.Gly336Cys

c.733G; p.Val245

W: 1%

AA: 30%

VSRH20c.[733C>G; 1006G>T]; p.[Leu245Val; Gly336Cys]

AA: 26-40%

All other populations: <0.01%

KellKKEL1KEL*01c.578C>T; p.Thr193Metc.578T; p.Met193

W: 9%

AA: 2%

A: Rare

Iranian Jews: 12%

Arabs: up to 25%

kKEL2KEL*02c.578C; p.Thr193

W: 99.8%

AA: 100%

KpaKEL3KEL*03c.841C>T; p.Arg281Trpc.841T; p.Trp281

W: 2%

AA: <0.01%

KpbKEL4KEL*04c.841C; p.Arg281All populations: 100%
JsaKEL6KEL*06c.1790T>C; p.Leu597Proc.1790C; p.Pro597

W: <0.01%

AA: 20%

JsbKEL7KEL*07c.1790T; p.Leu597

W: 100%

AA: 99%

DuffyFyaFY1FY*01c.125G>A; p.Gly42Aspc.125G; p.Gly42

W: 66%

AA: 10%

A: 99%

Thai: 97%

FybFY2FY*02c.125A; p.Asp42

W: 83%

AA: 23%

A: 18.5%

Thai: 31%

Chinese: 9.2%

Fyb-67C (Fyb ES)GATA FY-2FY*02N.01c.-67T>Cc.-67T>C

W: rare

AA: 68%

A: rare

FyxFY2WFY*02Mc.265C>T; p.Arg89Cysc.265C>T; p.Arg89Cysrare
KiddJkaJK1JK*01c.838G>A; p.Asp280Asnc.838G; p.Asp280

W: 77%

AA: 92%

A: 72%

JkbJK2JK*02c.838A; p.Asn280

W: 74%

AA: 49%

A: 76%

MNSMMNS1GYPA*01c.59T>C; p.Leu20Serc.59C; p.Ser20

W: 78%

A: 74%

NMNS2GYPA*02c.59T; p.Leu20

W: 72%

AA: 75%

SMNS3GYPB*03c.143C>T; p.Thr48Metc.143T; p.Met48

W: 55%

AA: 31%

sMNS4GYPB*04c.143C; p.Thr48

W: 89%

AA: 93%

UMNS5   

W: 99.9%

AA: 99%

Silencing S (Uvar)MNS-3,w5

GYPB*03N.01

GYPB*03N.02

GYPB*03N.03

GYPB*03N.04

c.230C>T; p.Thr77Met

or

c.270+5G>T

c.230C>T; p.Thr77Met

or

c.270+5G>T

rare
Silencing S (Uneg)MNS-3,-4,-5Deletion of GYPB exons 2-6 rare
LutheranLuaLU1LU*01c.230G>A; p.Arg77Hisc.230A; p.His77

W: 8%

AA: 5%

LubLU2LU*02c.230G; p.Arg77All populations: 99.8%
DombrockDoaDO1DO*01c. 793G>A; p.Asp265Asnc.793A; p.Asn265

W: 67%

AA: 55%

Japanese: 24%

Thai: 14%

DobDO2DO*02c.793G; p.Asp265

W: 82%

AA: 89%

HyDO4DO*04c.323G>T; p.Gly108Valc.323G; p.Gly108

AA: >99%

Most populations: 100%

 DO-4 c.323T; p.Val108rare
JoaDO5DO*05c.350C>T; p.Thr117Ilec.350C; p.Thr117

AA: >99%

Most populations: 100%

 DO-5 c.350T; p.Ile117rare
Landsteiner-Wiener (LW)LWaLW5LW*05c.299A>G; p.Gln100Argc.299G; p.Arg100All populations: 100%
LWbLW7LW*07c.299A; p.Gln100

Most populations: Rare

Estonians: 8%

Finns: 6%

Latvians and Lithuanians: 5%

Poles and Russians: 2%

Other Europeans:<1%

DiegoDiaDI1DI*01c.2561C>T; p.Pro854Leuc.2561T; p.Leu854

Most populations: 0.01%

South American Indians: 2% -54%

Japanese: 12%

Chinese: 5%

Hispanics: 1%

Poles: 0.47%

DibDI2DI*02c.2561C; p.Pro854

Most populations: 100%

Native Americans: 99%

ColtonCoaCO1CO*01c.134C>T; p.Ala45Valc.134C; p.Ala45All populations: 99.5%
CobCO2CO*02c.134T, p.Val45All populations: 10%
SciannaSc1SC1SC*01c.169G>A; p.Gly57Argc.169G; p.Gly57All populations: >99%
Sc2SC2SC*02c.169A; p.Arg57Europeans: 1%; more common in Mennonites
Hemoglobin SHbS  HBB c.20A>T; Glu6Valc.20A>T; Glu6ValAA: 10%

Reid, 2012 ; Dean, 2005 

A, Asian; AA, African American; ISBT, International Society for Blood Transfusion (ISBT) Committee on Terminology for Red Cell Surface Antigens; W, White

Reference transcripts: RHCE (NM_020485.5), KEL (NM_000420.2), ACKR1 (NM_002036.3; alt reference NM_001122951.2), SLC14A1 (NM_015865.6), GYPA (NM_002099.7), GYPB (NM_002100.5), BCAM (NM_005581.4), ART4 (NM_021071.2), ICAM4 (NM_001544.4), SLC4A1 (NM_000342.3), AQP1 (NM_198098.3), ERMAP (NM_018538.3)

References