Molecular analysis of immunized Jr(a-) or Lan- patients and validation of a high-throughput genotyping assay to screen blood donors for Jr(a-) and Lan- phenotypes

Transfusion. 2014 Jul;54(7):1836-46. doi: 10.1111/trf.12544. Epub 2014 Jan 24.

Abstract

Background: Individuals with anti-Jr(a) or anti-Lan are ideally transfused with rare Jr(a-) or Lan- red blood cells. We characterized mutations in Dutch Jr(a-) and Lan- individuals and developed a high-throughput genotyping assay to detect Jr(a-) and Lan- donors.

Study design and methods: Six Jr(a-) and seven Lan- persons, who all made anti-Jr(a) or anti-Lan, were sequenced for ABCG2 or ABCB6 and the copy number of ABCG2 and ABCB6 was determined. A total of 3366 Caucasian, 621 black, and 333 Chinese donors were screened with a high-throughput screening assay targeting frequently occurring mutations causing the Jr(a-) or Lan- phenotype.

Results: In the six tested Jr(a-) individuals previously described, c.376C > T, c.706C > T, and c.736C > T nonsense mutations in ABCG2 were detected. In the seven Lan- individuals 12 different mutations, of which 10 underlie the Lan- phenotype, were detected. No copy number variation was detected for ABCG2 and ABCB6. The high-throughput screening assay detected five Caucasian donors heterozygous for the c.706C > T or 736C > T mutation in ABCG2 and nine Caucasian donors heterozygous for the 574C > T mutation in ABCB6. No black or Chinese donors were found positive for a mutation.

Conclusion: We describe eight new mutations in ABCB6 of which seven, including three missense mutations, underlie the Lan- phenotype and determine that a complete gene deletion of ABCG2 or ABCB6 is not responsible for the Jr(a-) or Lan- phenotype, respectively. The extended heterogeneity of mutations causing the Jr(a-) or Lan- phenotype in most populations makes genetic screening for the Jr(a-) and Lan- phenotype inefficient in those populations.

Publication types

  • Validation Study

MeSH terms

  • ATP Binding Cassette Transporter, Subfamily G, Member 2
  • ATP-Binding Cassette Transporters / genetics
  • Alleles
  • Asian People / genetics
  • Base Sequence
  • Black People / genetics
  • Blood Donors*
  • Blood Group Antigens / genetics*
  • Blood Group Antigens / immunology*
  • Donor Selection
  • Genotyping Techniques*
  • High-Throughput Screening Assays / methods*
  • Humans
  • Immunization
  • Isoantigens / genetics
  • Isoantigens / immunology*
  • Molecular Sequence Data
  • Mutation
  • Neoplasm Proteins / genetics
  • Netherlands
  • Sequence Analysis, DNA / methods*
  • South Africa
  • White People / genetics

Substances

  • ABCB6 protein, human
  • ABCG2 protein, human
  • ATP Binding Cassette Transporter, Subfamily G, Member 2
  • ATP-Binding Cassette Transporters
  • Blood Group Antigens
  • Isoantigens
  • LAN antigen
  • Neoplasm Proteins

Associated data

  • GENBANK/AB829902
  • GENBANK/AB829903
  • GENBANK/AB829904
  • GENBANK/AB829905
  • GENBANK/AB829906
  • GENBANK/AB829907
  • GENBANK/AB854110
  • GENBANK/AB854111
  • GENBANK/KF831582
  • GENBANK/KF831583
  • GENBANK/KF831584
  • GENBANK/KF831585
  • GENBANK/KF831586
  • GENBANK/KF831587
  • GENBANK/KF831588
  • GENBANK/KF831589