Year-end Sale % OFF 
Abstract
Hereditary spherocytosis (HS) is a common anemia caused by germline mutations in red blood cell cytoskeleton proteins. The flow cytometry-based eosin-5′-maleimide (EMA) binding test is most frequently employed for reliable diagnostics. To perform this test, a number of healthy and ideally also age-matched controls are required, which can be challenging and complicates interlaboratory comparisons. To overcome this limitation, we modified the EMA binding test by replacing healthy controls with commercially available fluorescent beads. Blood samples from 289 individuals with suspected HS were analyzed using the EMA binding test with fluorescent beads and benchmarked against regular EMA binding test using two control samples. Using osmotic gradient ektacytometry as validation, 112 individuals (38.8%) were diagnosed with HS. Performance of the modified EMA binding test was not compromised (accuracy 90.3%) compared to EMA binding test using matched controls (accuracy 88.6%). Based on these findings, we conclude that the modified EMA binding test with fluorescent beads is an attractive alternative, especially in laboratories without easy access to matched controls. Furthermore, as fluorescent beads are stable and easily commutable, they could facilitate both interlaboratory comparisons and quality assessment programs.
Highlights
Hereditary spherocytosis (HS) is one of the most frequent hereditary hemolytic disorders in Caucasians, affecting approximately 1:2000
Blood samples from healthy controls were subjected to eosin-5 -maleimide (EMA) binding test with rainbow beads and osmotic gradient ektacytometry (Figure 1A)
We assessed whether commercially fluorescent beads can replace the need for numerous control samples in the EMA binding test with the aim of simplifying HS diagnostics
Summary
Hereditary spherocytosis (HS) is one of the most frequent hereditary hemolytic disorders in Caucasians, affecting approximately 1:2000. Affected individuals present with direct antiglobulin test (DAT) negative hemolytic anemia, increased mean corpuscular hemoglobin concentration (MCHC), and palpable splenomegaly, as well as a known family history of hemolytic anemia and gallbladder disease. The laboratory diagnosis of HS was based on the manual osmotic fragility (OF) test. This obsolete test measures the degree of hemolysis after exposing red blood cells (RBCs) to a salt solution of diminishing tonicity, a procedure that is laborious. Sensitivity and specificity of the Fluorescent Beads for EMA-Binding Test
Sign-in/Register to view highlights & summary 
Published Version (
Free)
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
