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Feasibility for non-invasive prenatal fetal blood group and platelet genotyping by massively parallel sequencing : A single test system for multiple atypical red cell, platelet and quality control markers

McGowan, Eunike C. LU orcid ; O'Brien, Helen ; Sarri, Mia E. ; Lopez, Genghis H. ; Daly, James J. ; Flower, Robert L. ; Gardener, Glenn J. and Hyland, Catherine A. (2024) In British Journal of Haematology 204(2). p.694-705
Abstract

Non-invasive prenatal tests (NIPT) to predict fetal red cell or platelet antigen status for alloimmunised women are provided for select antigens. This study reports on massively parallel sequencing (MPS) using a red cell and platelet probe panel targeting multiple nucleotide variants, plus individual identification single nucleotide polymorphisms (IISNPs). Maternal blood samples were provided from 33 alloimmunised cases, including seven with two red cell antibodies. Cell-free and genomic DNA was sequenced using targeted MPS and bioinformatically analysed using low-frequency variant detection. The resulting maternal genomic DNA allele frequency was subtracted from the cell-free DNA counterpart. Outcomes were matched against validated... (More)

Non-invasive prenatal tests (NIPT) to predict fetal red cell or platelet antigen status for alloimmunised women are provided for select antigens. This study reports on massively parallel sequencing (MPS) using a red cell and platelet probe panel targeting multiple nucleotide variants, plus individual identification single nucleotide polymorphisms (IISNPs). Maternal blood samples were provided from 33 alloimmunised cases, including seven with two red cell antibodies. Cell-free and genomic DNA was sequenced using targeted MPS and bioinformatically analysed using low-frequency variant detection. The resulting maternal genomic DNA allele frequency was subtracted from the cell-free DNA counterpart. Outcomes were matched against validated phenotyping/genotyping methods, where available. A 2.5% subtractive allele frequency threshold was set after comparing MPS predictions for K, RhC/c, RhE/e and Fya/Fyb against expected outcomes. This threshold was used for subsequent predictions, including HPA-15a, Jka/Jkb, Kpa/Kpb and Lua. MPS outcomes were 97.2% concordant with validated methods; one RhC case was discordantly negative and lacked IISNPs. IISNPs were informative for 30/33 cases as controls. NIPT MPS is feasible for fetal blood group genotyping and covers multiple blood groups and control targets in a single test. Noting caution for the Rh system, this has the potential to provide a personalised service for alloimmunised women.

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author
; ; ; ; ; ; and
publishing date
type
Contribution to journal
publication status
published
subject
keywords
alloimmunisation, blood groups, cell-free DNA, NIPT, sequencing
in
British Journal of Haematology
volume
204
issue
2
pages
694 - 705
publisher
John Wiley & Sons Inc.
external identifiers
  • scopus:85177445519
  • pmid:37984869
ISSN
0007-1048
DOI
10.1111/bjh.19197
language
English
LU publication?
no
additional info
Publisher Copyright: © 2023 The Authors. British Journal of Haematology published by British Society for Haematology and John Wiley & Sons Ltd.
id
23fd828d-c6dd-4575-abf6-f86d31df5041
date added to LUP
2024-12-20 10:07:23
date last changed
2025-06-06 23:25:14
@article{23fd828d-c6dd-4575-abf6-f86d31df5041,
  abstract     = {{<p>Non-invasive prenatal tests (NIPT) to predict fetal red cell or platelet antigen status for alloimmunised women are provided for select antigens. This study reports on massively parallel sequencing (MPS) using a red cell and platelet probe panel targeting multiple nucleotide variants, plus individual identification single nucleotide polymorphisms (IISNPs). Maternal blood samples were provided from 33 alloimmunised cases, including seven with two red cell antibodies. Cell-free and genomic DNA was sequenced using targeted MPS and bioinformatically analysed using low-frequency variant detection. The resulting maternal genomic DNA allele frequency was subtracted from the cell-free DNA counterpart. Outcomes were matched against validated phenotyping/genotyping methods, where available. A 2.5% subtractive allele frequency threshold was set after comparing MPS predictions for K, RhC/c, RhE/e and Fy<sup>a</sup>/Fy<sup>b</sup> against expected outcomes. This threshold was used for subsequent predictions, including HPA-15a, Jk<sup>a</sup>/Jk<sup>b</sup>, Kp<sup>a</sup>/Kp<sup>b</sup> and Lu<sup>a</sup>. MPS outcomes were 97.2% concordant with validated methods; one RhC case was discordantly negative and lacked IISNPs. IISNPs were informative for 30/33 cases as controls. NIPT MPS is feasible for fetal blood group genotyping and covers multiple blood groups and control targets in a single test. Noting caution for the Rh system, this has the potential to provide a personalised service for alloimmunised women.</p>}},
  author       = {{McGowan, Eunike C. and O'Brien, Helen and Sarri, Mia E. and Lopez, Genghis H. and Daly, James J. and Flower, Robert L. and Gardener, Glenn J. and Hyland, Catherine A.}},
  issn         = {{0007-1048}},
  keywords     = {{alloimmunisation; blood groups; cell-free DNA; NIPT; sequencing}},
  language     = {{eng}},
  number       = {{2}},
  pages        = {{694--705}},
  publisher    = {{John Wiley & Sons Inc.}},
  series       = {{British Journal of Haematology}},
  title        = {{Feasibility for non-invasive prenatal fetal blood group and platelet genotyping by massively parallel sequencing : A single test system for multiple atypical red cell, platelet and quality control markers}},
  url          = {{http://dx.doi.org/10.1111/bjh.19197}},
  doi          = {{10.1111/bjh.19197}},
  volume       = {{204}},
  year         = {{2024}},
}