Delayed Measurement of Eosin-5′-Maleimide Binding May Affect the Test Results of Highly Hemolyzed Samples In Vivo and In Vitro—A Case Study

Ciepiela, Olga; Adamowicz-Salach, Anna; Zdziechowicz, Izabela; Kotuła, Iwona

doi:10.1097/mph.0000000000000652

Cited by 3 publications

(1 citation statement)

References 9 publications

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“…The laboratory diagnosis of HS mainly relies on red blood cell (RBC) morphology examination, osmotic fragility (OF) test, acidized glycerolysis test (AGLT), and maleimide eosin (EMA) ( Farias, 2017 ). Although the EMA test showed maximum disease specificity, no single test can identify all cases of hereditary spherocytosis ( Bianchi et al, 2012 ; Ciepiela et al, 2016 ). At present, molecular tests based on next-generation sequencing technology are widely used in the differential diagnosis and confirmation of pathogenic variants, further improving the accuracy and specificity of the diagnosis of HS ( Xue et al, 2019 ; Richmond et al, 2020 ; Xue et al, 2020 ; Fan et al, 2021 ; Xie et al, 2021 ).…”

Section: Introductionmentioning

confidence: 99%

Integrative preimplantation genetic testing analysis for a Chinese family with hereditary spherocytosis caused by a novel splicing variant of SPTB

Tian,

Wang,

Yang

et al. 2023

Front. Genet.

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Hereditary spherocytosis (HS), the most common inherited hemolytic anemia disorder, is characterized by osmotically fragile microspherocytic red cells with a reduced surface area on the peripheral blood smear. Pathogenic variants in five erythrocyte membrane structure-related genes ANK1 (Spherocytosis, type 1; MIM#182900), SPTB (Spherocytosis, type 2; MIM#616649), SPTA1 (Spherocytosis, type 3; MIM#270970), SLC4A1 (Spherocytosis, type 4; MIM#612653) and EPB42 (Spherocytosis, type 5; MIM#612690) have been confirmed to be related to HS. There have been many studies on the pathogenic variants and mechanisms of HS, however, studies on how to manage the transmission of HS to the next-generation have not been reported. In this study, we recruited a patient with HS. Targeted next-generation sequencing with a panel of 208 genes related to blood system diseases detected a novel heterozygous variant in the SPTB: c.300+2dup in the proband. Sanger sequencing of variant alleles and haplotype linkage analysis of single nucleotide polymorphism (SNP) based on next-generation sequencing were performed simultaneously. Five embryos were identified with one heterozygous and four not carrying the SPTB variant. Single-cell amplification and whole genome sequencing showed that three embryos had varying degrees of trisomy mosaicism. One of two normal embryos was transferred to the proband. Ultimately, a healthy boy was born, confirmed by noninvasive prenatal testing for monogenic conditions (NIPT-M) to be disease-free. This confirmed our successful application of PGT in preventing transmission of the pathogenic variant allele in the HS family.

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Section: Introductionmentioning

confidence: 99%

Integrative preimplantation genetic testing analysis for a Chinese family with hereditary spherocytosis caused by a novel splicing variant of SPTB

Tian,

Wang,

Yang

et al. 2023

Front. Genet.

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Overview on Hereditary Spherocytosis Diagnosis

Polizzi,

Dicembre,

Failla

et al. 2024

Int J Lab Hematology

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IntroductionHereditary spherocytosis (HS) is a congenital haemolytic disorder, resulting from plasma membrane protein deficiency of red blood cells (RBCs). Typical pathological signs are anemia, jaundice, and splenomegaly; in newborns, jaundice is the main symptom.Material and MethodsThis study focused on the state of art about the HS diagnosis, from traditional to innovative methods, including diagnostic algorithms that can be applied for pediatric and adult patients, for different laboratory diagnostic levels.ResultsThe first erythrocyte parameters used for HS diagnosis were the mean corpuscular hemoglobin concentration (MCHC), mean corpuscular volume (MCV), and red blood cell distribution width (RDW); nowadays new parameters are used in blood cell counter. Advia analyzers (Siemens Medical Solutions) supply the hyper‐dense cell percentage (% Hyper), which reflects the red blood cells hyperchromia. Sysmex instruments (i.e. XT‐4000i, XE‐5000, XN‐Series) provide the MicroR, that is the percentage of erythrocytes smaller than 60 fL, Hypo‐He, which is the percentage of erythrocytes with a content of hemoglobin less than 17 pg and % Hyper‐He, which represents the percentage of RBC with cellular hemoglobin content higher than 49 pg. CELL‐DYN Sapphire (Abbott Diagnostics) introduced the HPR parameter (% HPR), which represents the erythrocytes with hemoglobin > 410 g/L. Beckman Coulter instruments supply the mean sphered corpuscular volume (MSCV), which is the average volume of all erythrocytes, including mature erythrocytes and reticulocytes. Other reference tests for screening and diagnosis of HS are the acidified glycerol lysis test (AGLT), the eosin‐5‐maleimide (EMA) binding test and genetic testing by next‐generation sequencing.ConclusionsThe diagnostic workup of hereditary spherocytosis could be improved thanks to all the available tests, including new molecular tools. However, it requires synergy between clinicians and laboratory staff, evaluating clinical manifestations, all available data related to the disease and the prognosis to fill the diagnostic gaps in the near future.

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Advances in laboratory diagnosis of hereditary spherocytosis

Farias

2017

Clinical Chemistry and Laboratory Medicine (CCLM)

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Among the red cell membrane disorders, hereditary spherocytosis (HS) is one of the most common causes of inherited hemolytic anemia. HS results from the deficiency or dysfunction of red blood cell membrane proteins, such as α spectrin, β spectrin, ankyrin, anion channel protein (Band-3 protein), protein 4.1 and protein 4.2. Conventionally, HS diagnosis is established through a series of tests, which include spherocytes identification in peripheral smear, reticulocyte count, osmotic fragility, etc. Currently, different hematological analyzers provide erythrocyte indicators that estimate the presence of spherocytes and correlate that with HS, which can be useful for disease screening. The most traditional method is the osmotic fragility (OF) test, which is labor-intensive and time-consuming to perform and presents low sensitivity and specificity values. Thus, new methods have been developed for HS diagnosis, such as flow cytometry. Current guidelines recommend the use of flow cytometry as a screening test for HS diagnosis using the eosin-5'-maleimide (EMA) binding test. Thus, HS diagnosis is the result of a collaboration between clinicians and laboratories, who should take into account the family history and the exclusion of other causes of secondary spherocytosis.

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Delayed Measurement of Eosin-5′-Maleimide Binding May Affect the Test Results of Highly Hemolyzed Samples In Vivo and In Vitro—A Case Study

Cited by 3 publications

References 9 publications

Integrative preimplantation genetic testing analysis for a Chinese family with hereditary spherocytosis caused by a novel splicing variant of SPTB

Integrative preimplantation genetic testing analysis for a Chinese family with hereditary spherocytosis caused by a novel splicing variant of SPTB

Overview on Hereditary Spherocytosis Diagnosis

Advances in laboratory diagnosis of hereditary spherocytosis

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