Real-world performance of a clinical droplet digital polymerase chain reaction assay for non-invasive foetal blood group and platelet antigen genotyping of alloimmunized pregnant women with antibodies directed against RhD, RhE, Rhc, RhC, K1, HPA-1a or HPA-5b: A 1-year experience.

IF 1.8 4区医学 Q3 HEMATOLOGY Vox Sanguinis Pub Date : 2024-12-08 DOI:10.1111/vox.13777

Camilla Calandrini, Onno J H M Verhagen, Ahmed Tissoudali, Christa H E Homburg, Jessica Vessies, Mark Brussee, Erik H van Beers, C Ellen van der Schoot, Masja de Haas

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Abstract

Background and objectives: To test the performance of a new droplet digital polymerase chain reaction (ddPCR) non-invasive foetal blood group and platelet antigen genotyping assay in the setting of a Dutch reference laboratory for foetal blood group and platelet antigen genotyping. Our population comprised 229 consecutive alloimmunized pregnant women who presented between April 2022 and March 2023 with 250 requests for non-invasive foetal RHD, RHE, RHc, RHC, K1, HPA-1a or HPA-5b blood group and platelet antigen genotyping.

Materials and methods: Samples were genotyped for blood group and platelet antigen alleles along with methylated RASSF1a (mRASSF1a) and sex-determining region of Y (SRY) and DYS14 as positive foetal controls. Negative blood group and platelet antigen results were issued only when foetal controls were positive; otherwise, such samples were classified as inconclusive.

Results: The assay achieved a success rate of 98.4% (246 of 250) because one case was lost to follow-up, one case was solved with quantitative polymerase chain reaction (qPCR) and one case precluded foetal typing due to RHD variant mothers. Only 10 cases needed a second sample and one case a third for a valid final result. We identified 116 maternal-foetal blood group and platelet antigen incompatibilities.

Conclusion: Clinical non-invasive foetal blood group and platelet antigen typing of alloimmunized pregnant women via ddPCR is successful and represents an improvement over qPCR because of the addition of a foetal control and because ddPCR circumvents potential interference from maternal cell-free DNA (cfDNA) background for foetal HPA-1 and K1.

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使用针对RhD、RhE、Rhc、Rhc、K1、HPA-1a或HPA-5b的抗体进行同种异体免疫孕妇的无创胎儿血型和血小板抗原基因分型的临床液滴数字聚合酶链反应试验的实际表现：1年的经验。

背景与目的：在荷兰某胎儿血型与血小板抗原基因分型参比实验室中，检测新型液滴数字聚合酶链反应（ddPCR）无创胎儿血型与血小板抗原基因分型检测方法的性能。我们的人群包括229名在2022年4月至2023年3月期间连续接受同种免疫的孕妇，其中250名要求进行非侵入性胎儿RHD、RHE、RHc、RHc、K1、HPA-1a或HPA-5b血型和血小板抗原基因分型。材料和方法：对样本进行血型和血小板抗原等位基因的基因分型，并将甲基化的RASSF1a （mRASSF1a）和Y基因性别决定区（SRY）和DYS14作为阳性胎儿对照。阴性血型和血小板抗原结果只有在胎儿对照阳性时才会公布；否则，这些样本被归类为不确定。结果：1例失访，1例用定量聚合酶链反应（qPCR）解决，1例因RHD变异母亲而排除胎儿分型，检测成功率为98.4%（246 / 250）。只有10例需要第二次采样，1例需要第三次采样才能获得有效的最终结果。我们鉴定了116例母胎血型和血小板抗原不相容。结论：通过ddPCR进行同种异体免疫孕妇的临床无创胎儿血型和血小板抗原分型是成功的，并且由于ddPCR增加了胎儿对照，并且由于ddPCR规避了母体游离DNA （cfDNA）背景对胎儿HPA-1和K1的潜在干扰，因此比qPCR有改进。

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来源期刊

Vox Sanguinis 医学-血液学

CiteScore

4.40

自引率

11.10%

发文量

156

审稿时长

6-12 weeks

期刊介绍： Vox Sanguinis reports on important, novel developments in transfusion medicine. Original papers, reviews and international fora are published on all aspects of blood transfusion and tissue transplantation, comprising five main sections: 1) Transfusion - Transmitted Disease and its Prevention: Identification and epidemiology of infectious agents transmissible by blood; Bacterial contamination of blood components; Donor recruitment and selection methods; Pathogen inactivation. 2) Blood Component Collection and Production: Blood collection methods and devices (including apheresis); Plasma fractionation techniques and plasma derivatives; Preparation of labile blood components; Inventory management; Hematopoietic progenitor cell collection and storage; Collection and storage of tissues; Quality management and good manufacturing practice; Automation and information technology. 3) Transfusion Medicine and New Therapies: Transfusion thresholds and audits; Haemovigilance; Clinical trials regarding appropriate haemotherapy; Non-infectious adverse affects of transfusion; Therapeutic apheresis; Support of transplant patients; Gene therapy and immunotherapy. 4) Immunohaematology and Immunogenetics: Autoimmunity in haematology; Alloimmunity of blood; Pre-transfusion testing; Immunodiagnostics; Immunobiology; Complement in immunohaematology; Blood typing reagents; Genetic markers of blood cells and serum proteins: polymorphisms and function; Genetic markers and disease; Parentage testing and forensic immunohaematology. 5) Cellular Therapy: Cell-based therapies; Stem cell sources; Stem cell processing and storage; Stem cell products; Stem cell plasticity; Regenerative medicine with cells; Cellular immunotherapy; Molecular therapy; Gene therapy.