Glenda M. Millard

Evaluation of targeted exome sequencing for 28 protein‐based blood group systems, including the homologous gene systems, for blood group genotyping

Blood group single nucleotide polymorphism genotyping probes for a limited range of polymorphisms. This study investigated whether massively parallel sequencing (also known as next‐generation sequencing), with a targeted exome strategy, provides an extended blood group genotype and the extent to which massively parallel sequencing correctly genotypes in homologous gene systems, such as RH and MNS.

Noninvasive fetal RHD genotyping by microfluidics digital PCR using maternal plasma from two alloimmunized women with the variant RHD(IVS3+1G>A) allele

Whats already known about this topic? Noninvasive prenatal RHD typing can be achieved by using cell-free fetal DNA in the plasma of RhD-negative mothers. For RhD-negative mothers carrying intact but dysfunctional RHD gene variants, the abundant maternal RHD sequences in maternal plasma could interfere with the fetal RHD allele detection. What does this study add? Digital PCR provides a high analytical specificity to noninvasively determine the fetal inheritance of RHD allele in alloimmunized pregnancies involving maternal RHD variants.

A D+ blood donor with a novel RHD*D‐CE(5‐6)‐D gene variant exhibits the low‐frequency antigen RH23 (DW) characteristic of the partial DVa phenotype

Blood donors whose red blood cells (RBCs) exhibit a partial RhD phenotype, lacking some D epitopes, present as D+ in routine screening. Such phenotypes can exhibit low‐frequency antigens (LFAs) of clinical significance. The aim of this study was to describe the serologic and genetic profile for a blood donor with an apparent D+ phenotype carrying a variant RHD gene where D Exons 5 and 6 are replaced by RHCE Exon (5‐6).

Targeted exome sequencing defines novel and rare variants in complex blood group serology cases for a red blood cell reference laboratory setting

We previously demonstrated that targeted exome sequencing accurately defined blood group genotypes for reference panel samples characterized by serology and single‐nucleotide polymorphism (SNP) genotyping. Here we investigate the application of this approach to resolve problematic serology and SNP‐typing cases.

Strategy for managing maternal variant RHD alleles in Rhesus D negative obstetric populations during fetal RHD genotyping

Fetal RHD screening programs that aim to reduce unnecessary antenatal anti‐D prophylaxis are being introduced into clinical practice. Strategies to manage women serologically typed as Rhesus D negative who have maternal RHD variants are needed. This study describes maternal RHD allelic variants detected in nonselected and alloimmunised Rhesus D negative obstetric populations and explores a mathematical approach to identify these variants.

A proposed new low-frequency antigen in the Augustine blood group system associated with a severe case of hemolytic disease of the fetus and newborn: Proposed new low-frequency antigen in the Aug Blood Group System

A male infant born at 29 weeks presented with a severe case of hemolytic disease of the fetus and newborn (HDFN) (cord blood direct antiglobulin test 41, hemoglobin 45 g/L with cardiac failure, pleural effusion, and generalized edema). Two exchange and four top-up transfusions were required. The maternal antibody was reactive with paternal red blood cells (RBCs). Later testing revealed that the antibody reacted with RBCs from four additional members of the paternal family. Extensive testing excluded clinically relevant RBC antibodies but failed to reveal a specificity for this antibody. To guide further investigation, specimens from the family (n 5 10) were submitted for blood group genetic studies.

Genotyping by sequencing defines independent novel RHD variants for an antenatal patient and a blood donor

A ccurate D blood group phenotyping is important in transfusion medicine to guide antenatal management for D2 women and donation management for red blood cells (RBCs) from blood donors. A small percentage of individuals carry variant RHD genes associated with either partial D, weak D, or the DEL phenotype, and the latter are detected only by laborious adsorption and elution techniques. Single nucleotide polymorphism (SNP)-microarray typing detects such variant RHD genes but is limited in the number of polymorphisms it interrogates. This report describes the resolution by DNA sequencing of two samples that were unresolved by SNP-microarray genotyping: an antenatal sample (S1) from a patient who presented with variable D reactivity patterns, and a blood donor sample (S2) from a donor who presented as D2 on routine serology but had an E1 phenotype, which sometimes is associated with a DEL phenotype.

Noninvasive fetal RHD genotyping of RhD negative pregnant women for targeted anti‐D therapy in Australia: A cost‐effectiveness analysis

To undertake a cost‐effectiveness analysis of noninvasive fetal RHD genotyping to target pregnant women for antenatal anti‐D prophylaxis therapy.

A DEL phenotype attributed to RHD Exon 9 sequence deletion: slipped-strand mispairing and blood group polymorphisms: SLIPPED-STRAND MISPAIRING MECHANISM IN RHD

The RhD blood group antigen is extremely polymorphic and the DEL phenotype represents one such class of polymorphisms. The DEL phenotype prevalent in East Asian populations arises from a synonymous substitution defined as RHD*1227A. However, initially, based on genomic and cDNA studies, the genetic basis for a DEL phenotype in Taiwan was attributed to a deletion of RHD Exon 9 that was never verified at the genomic level by any other independent group. Here we investigate the genetic basis for a Caucasian donor with a DEL partial D phenotype and compare the genomic findings to those initial molecular studies.

Investigation of the variable In(Lu) phenotype caused by KLF1 variants: THE In(Lu) PHENOTYPE AND KLF1 VARIANTS

KLF1 is an essential transcriptional activator that drives erythropoiesis. KLF1 variants can result in the Inhibitor of Lutheran, or In(Lu), phenotype where red blood cells (RBCs) have reduced BCAM (LU) and CD44 (IN). Other RBC surface molecules also have changed expression; however, there is controversy in the literature regarding which are truly impacted. We aimed to investigate KLF1 variants in the Australian population.