Available online 6 February 2024, 113628
Author links open overlay panel, , AbstractThe importance of structural genetic variants, such as copy number variations (CNVs), in modulating human disease is being increasingly recognized. Several clinical conditions require investigation of human neutrophil antigen (HNA-1), which is encoded by the Fc gamma receptor IIIb gene (FCGR3B), including suspicion of neutropenia, infections, and proactive testing of blood component donors to reduce the potential risk in transfusion. In this study, we compared real-time quantitative polymerase chain reaction (qPCR) with two digital PCR (dPCR) platforms, namely droplet digital PCR and an array-based platform, to determine CNV in FCGR3B. We initially tested 400 anonymous blood donors with qPCR using a commercially available TaqMan probe assay (Applied Biosystems) on a Quant Studio 12 Flex. CNV was determined for all 400 tested individuals with copy numbers ranging from zero to four. Zero copies were detected in 0.2% (1/400), one copy was detected in 3.8% (15/400), two copies were detected in 87.8% (351/400), three copies were detected in 8.0% (32/400), and four copies were detected in 0.2% (1/400) of tested individuals. From this cohort, we selected 32 donors with copy numbers from zero to four for analyses with Digital Real-Time PCR (dPCR) using Lab on an array (LOAA) on an On-Point analyzer from Optolane Technologies Inc. and the Droplet Digital PCR (ddPCR) platform from Bio-Rad Laboratories. We compared the obtained copy numbers of FCGR3B on the three platforms and found full concordance between the copy numbers obtained. We therefore conclude that all three platforms can be used for quantification of copy numbers for FCGR3B, and although dPCR has some advantages over qPCR, it was not necessary for reliably estimating CNV of the FCGR3B gene.
KeywordsFCGR3B
CNV
Digital PCR
Droplet digital PCR
qPCR
AbbreviationsdPCRdigital real-time PCR
HNAhuman neutrophil antigen
MLPAMultiplex ligation-dependent probe amplification
PCRpolymerase chain reaction
PCR-SSPsequence-specific primer PCR
qPCRreal-time quantitative PCR
SNPsingle nucleotide polymorphism.
© 2024 The Authors. Published by Elsevier B.V.
留言 (0)