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

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ABO genotyping with next-generation sequencing to resolve heterogeneity in donors with serology discrepancies. / Wu, Ping Chun; Lin, Yin-Hung; Tsai, Lei Fang; Chen, Ming Hung; Chen, Pei-Lung; Pai, Shun-Chung.

In: Transfusion, Vol. 58, No. 9, 09.2018, p. 2232-2242.

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Wu, Ping Chun ; Lin, Yin-Hung ; Tsai, Lei Fang ; Chen, Ming Hung ; Chen, Pei-Lung ; Pai, Shun-Chung. / ABO genotyping with next-generation sequencing to resolve heterogeneity in donors with serology discrepancies. In: Transfusion. 2018 ; Vol. 58, No. 9. pp. 2232-2242.

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TY - JOUR

T1 - ABO genotyping with next-generation sequencing to resolve heterogeneity in donors with serology discrepancies

AU - Wu, Ping Chun

AU - Lin, Yin-Hung

AU - Tsai, Lei Fang

AU - Chen, Ming Hung

AU - Chen, Pei-Lung

AU - Pai, Shun-Chung

N1 - © 2018 AABB.

PY - 2018/9

Y1 - 2018/9

N2 - 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.

AB - 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.

KW - ABO Blood-Group System/analysis

KW - Blood Donors

KW - Blood Group Incompatibility/diagnosis

KW - Blood Grouping and Crossmatching/methods

KW - Cell Separation

KW - Chimerism

KW - DNA/genetics

KW - Flow Cytometry

KW - Genotype

KW - Genotyping Techniques/methods

KW - High-Throughput Nucleotide Sequencing/methods

KW - Humans

KW - Introns/genetics

KW - Polymorphism, Single Nucleotide

KW - Reproducibility of Results

KW - Sequence Analysis, DNA/methods

KW - Sequence Deletion

KW - Serologic Tests

KW - Transfusion Reaction/prevention & control

U2 - 10.1111/trf.14654

DO - 10.1111/trf.14654

M3 - Article

C2 - 29770457

VL - 58

SP - 2232

EP - 2242

JO - Transfusion

JF - Transfusion

SN - 1537-2995

IS - 9

ER -