Prediction of fetal blood group antigens from maternal plasma using Ion AmpliSeq HD technology

Autor: Agnieszka Orzińska, Anna Kluska, Aneta Balabas, Magdalena Piatkowska, Maria Kulecka, Jerzy Ostrowski, Michal Mikula, Marzena Dębska, Małgorzata Uhrynowska, Katarzyna Guz
Rok vydání: 2022
Předmět:
Zdroj: Transfusion. 62:458-468
ISSN: 1537-2995
0041-1132
DOI: 10.1111/trf.16780
Popis: Fetal blood group (BG) and platelet (HPA) antigens may trigger maternal immunization, causing a fetal disease. Noninvasive prenatal diagnostics (NIPT) predicts fetal genotype, identifying pregnancies with no risk. All current techniques detect fetal antigen alleles with unspecific background and without estimation of fetal fraction, thus new protocols for detection of fetal BG/HPA alleles with ultrahigh sensitivity still need to be tested to improve NIPT.To design NIPT of clinically important antigens using Ion AmpliSeq HD technology.Plasma DNA from 36 pregnant women (9-33 week of gestation, 24 immunized with anti-HPA-1a,-3b,-15a, -K, or -D+C+S), with known BG/HPA genotypes of their neonates/partners, was tested on Ion S5 System using the Ion AmpliSeq HD designer custom gene panel. NGS contained 25 rs-targets encoding relevant BG/HPA antigens and 10 markers.Using the NGS protocol, 76 out of 85 differences in fetal/maternal BG/HPA genotypes were determined in concentration above 2% fetal paternally inherited allele chimerism. The level of unspecific reads for BG/HPA alleles was below 0.87%. In 24 immunized women NGS revealed feto-maternal incompatibility in 11 cases (from 2.44% to 7.41%) and excluded in 10 (0.05%), three cases had inconclusive results (1.79%, 0.19%, 0.11%). The presence of fetal DNA was confirmed in each case by detecting markers with at least 2% chimerism.The use of Ion AmpliSeq HD technology improves the prediction of feto-maternal incompatibility, increasing the sensitivity of BG/HPA NIPT and serving confirmation of the fetal DNA at the same workflow.
Databáze: OpenAIRE
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