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ABO genotyping with next-generation sequencing to resolve heterogeneity in donors with serology discrepancies

Wu, Ping Chun LU orcid ; Lin, Yin-Hung ; Tsai, Lei Fang ; Chen, Ming Hung ; Chen, Pei-Lung and Pai, Shun-Chung (2018) In Transfusion 58(9). p.2232-2242
Abstract

BACKGROUND: ABO subtypes are characterized by the alteration of antigens present and their expression levels on red blood cells and many are linked to genetic changes in the ABO gene. Weakened expression of antigens should be identified to prevent transfusion reactions or ABO-incompatible transplantations. Genotyping can be applied to identify subtypes to complement serologic testing. Next-generation sequencing (NGS) has shown to provide sensitive and accurate genotyping results as well as valuable cis/trans information. Here we took advantage of NGS and applied it to resolve serology discrepancies in ABO typing.

STUDY DESIGN AND METHODS: In this study, we customized capture probes targeting the entire ABO gene and sequenced on... (More)

BACKGROUND: ABO subtypes are characterized by the alteration of antigens present and their expression levels on red blood cells and many are linked to genetic changes in the ABO gene. Weakened expression of antigens should be identified to prevent transfusion reactions or ABO-incompatible transplantations. Genotyping can be applied to identify subtypes to complement serologic testing. Next-generation sequencing (NGS) has shown to provide sensitive and accurate genotyping results as well as valuable cis/trans information. Here we took advantage of NGS and applied it to resolve serology discrepancies in ABO typing.

STUDY DESIGN AND METHODS: In this study, we customized capture probes targeting the entire ABO gene and sequenced on MiSeq Illumina. The subtype-causing variants were identified, and cis/trans association to ABO alleles was determined. The results from NGS, serology, and Sanger sequencing were compared.

RESULTS: Four control samples typed A, B, O, and AB were correctly genotyped. Of 24 serologically discrepant samples, subtype-causing variations were found in 20 cases, with two unresolved and two identified as weakening of ABO antibody in reverse. The types of variations include 17 known subtype alleles, one novel variant, one novel large deletion, and one microchimerism. Haplotypes encompassing Exons 6 and 7 of ABO were reconstructed in 17 of the 20 samples.

CONCLUSION: This study demonstrated a full coverage of ABO by capture-based panel, phasing analysis with NGS in ABO genotyping resolved heterogeneity with novel allele and microchimerism findings. This approach provided a more precise method for subtyping and thereby leading to safer transfusion.

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author
; ; ; ; and
publishing date
type
Contribution to journal
publication status
published
subject
keywords
ABO Blood-Group System/analysis, Blood Donors, Blood Group Incompatibility/diagnosis, Blood Grouping and Crossmatching/methods, Cell Separation, Chimerism, DNA/genetics, Flow Cytometry, Genotype, Genotyping Techniques/methods, High-Throughput Nucleotide Sequencing/methods, Humans, Introns/genetics, Polymorphism, Single Nucleotide, Reproducibility of Results, Sequence Analysis, DNA/methods, Sequence Deletion, Serologic Tests, Transfusion Reaction/prevention & control
in
Transfusion
volume
58
issue
9
pages
11 pages
publisher
Wiley-Blackwell
external identifiers
  • pmid:29770457
  • scopus:85047454574
ISSN
1537-2995
DOI
10.1111/trf.14654
language
English
LU publication?
no
additional info
© 2018 AABB.
id
7d608983-65a3-4006-8f62-bada0f0f4cac
date added to LUP
2019-10-11 14:14:18
date last changed
2024-05-15 00:06:33
@article{7d608983-65a3-4006-8f62-bada0f0f4cac,
  abstract     = {{<p>BACKGROUND: ABO subtypes are characterized by the alteration of antigens present and their expression levels on red blood cells and many are linked to genetic changes in the ABO gene. Weakened expression of antigens should be identified to prevent transfusion reactions or ABO-incompatible transplantations. Genotyping can be applied to identify subtypes to complement serologic testing. Next-generation sequencing (NGS) has shown to provide sensitive and accurate genotyping results as well as valuable cis/trans information. Here we took advantage of NGS and applied it to resolve serology discrepancies in ABO typing.</p><p>STUDY DESIGN AND METHODS: In this study, we customized capture probes targeting the entire ABO gene and sequenced on MiSeq Illumina. The subtype-causing variants were identified, and cis/trans association to ABO alleles was determined. The results from NGS, serology, and Sanger sequencing were compared.</p><p>RESULTS: Four control samples typed A, B, O, and AB were correctly genotyped. Of 24 serologically discrepant samples, subtype-causing variations were found in 20 cases, with two unresolved and two identified as weakening of ABO antibody in reverse. The types of variations include 17 known subtype alleles, one novel variant, one novel large deletion, and one microchimerism. Haplotypes encompassing Exons 6 and 7 of ABO were reconstructed in 17 of the 20 samples.</p><p>CONCLUSION: This study demonstrated a full coverage of ABO by capture-based panel, phasing analysis with NGS in ABO genotyping resolved heterogeneity with novel allele and microchimerism findings. This approach provided a more precise method for subtyping and thereby leading to safer transfusion.</p>}},
  author       = {{Wu, Ping Chun and Lin, Yin-Hung and Tsai, Lei Fang and Chen, Ming Hung and Chen, Pei-Lung and Pai, Shun-Chung}},
  issn         = {{1537-2995}},
  keywords     = {{ABO Blood-Group System/analysis; Blood Donors; Blood Group Incompatibility/diagnosis; Blood Grouping and Crossmatching/methods; Cell Separation; Chimerism; DNA/genetics; Flow Cytometry; Genotype; Genotyping Techniques/methods; High-Throughput Nucleotide Sequencing/methods; Humans; Introns/genetics; Polymorphism, Single Nucleotide; Reproducibility of Results; Sequence Analysis, DNA/methods; Sequence Deletion; Serologic Tests; Transfusion Reaction/prevention & control}},
  language     = {{eng}},
  number       = {{9}},
  pages        = {{2232--2242}},
  publisher    = {{Wiley-Blackwell}},
  series       = {{Transfusion}},
  title        = {{ABO genotyping with next-generation sequencing to resolve heterogeneity in donors with serology discrepancies}},
  url          = {{http://dx.doi.org/10.1111/trf.14654}},
  doi          = {{10.1111/trf.14654}},
  volume       = {{58}},
  year         = {{2018}},
}