{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,24]],"date-time":"2026-06-24T16:01:14Z","timestamp":1782316874308,"version":"3.54.5"},"reference-count":31,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2023,2,17]],"date-time":"2023-02-17T00:00:00Z","timestamp":1676592000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"National Research Foundation of Korea (NRF) grants funded by the Korean government (MSIT; Ministry of Science and ICT)","award":["2020R1F1A1077316"],"award-info":[{"award-number":["2020R1F1A1077316"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Diagnostics"],"abstract":"<jats:p>Background: Hereditary hemolytic anemia (HHA) is defined as a group of heterogeneous and rare diseases caused by defects of red blood cell (RBC) metabolism and RBC membrane, which leads to lysis or premature clearance. The aim of this study was to investigate individuals with HHA for potential disease-causing variants in 33 genes reported to be associated with HHA. Methods: A total of 14 independent individuals or families diagnosed with suspected HHA, and in particular, RBC membranopathy, RBC enzymopathy, and hemoglobinopathy, were collected after routine peripheral blood smear testing. A custom designed panel, including the 33 genes, was performed using gene panel sequencing on the Ion Torrent PGM\u2122 Dx System. The best candidate disease-causing variants were confirmed by Sanger sequencing. Results: Several variants of the HHA-associated genes were detected in 10 out of 14 suspected HHA individuals. After excluding those variants predicted to be benign, 10 pathogenic variants and 1 variant of uncertain significance (VUS) were confirmed in 10 individuals with suspected HHA. Of these variants, the p.Trp704Ter nonsense variant of EPB41 and missense p.Gly151Asp variant of SPTA1 were identified in two out of four hereditary elliptocytoses. The frameshift p.Leu884GlyfsTer27 variant of ANK1, nonsense p.Trp652Ter variant of the SPTB, and missense p.Arg490Trp variant of PKLR were detected in all four hereditary spherocytosis cases. Missense p.Glu27Lys, nonsense p.Lys18Ter variants, and splicing errors such as c.92 + 1G &gt; T and c.315 + 1G &gt; A within HBB were identified in four beta thalassemia cases. Conclusions: This study provides a snapshot of the genetic alterations in a cohort of Korean HHA individuals and demonstrates the clinical utility of using gene panels in HHA. Genetic results can provide precise clinical diagnosis and guidance regarding medical treatment and management for some individuals.<\/jats:p>","DOI":"10.3390\/diagnostics13040770","type":"journal-article","created":{"date-parts":[[2023,2,20]],"date-time":"2023-02-20T05:57:52Z","timestamp":1676872672000},"page":"770","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Five Years\u2019 Experience with Gene Panel Sequencing in Hereditary Hemolytic Anemia Screened by Routine Peripheral Blood Smear Examination"],"prefix":"10.3390","volume":"13","author":[{"given":"Namsu","family":"Kim","sequence":"first","affiliation":[{"name":"Department of Laboratory Medicine, Jeonbuk National University Medical School and Hospital, Jeonju 54907, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tae Yun","family":"Kim","sequence":"additional","affiliation":[{"name":"Department of Thoracic and Cardiovascular Surgery, Jeonbuk National University Medical School and Hospital, Jeonju 54907, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4174-5266","authenticated-orcid":false,"given":"Ji Yoon","family":"Han","sequence":"additional","affiliation":[{"name":"Department of Pediatrics, College of Medicine, The Catholic University of Korea, Seoul 06591, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7354-4234","authenticated-orcid":false,"given":"Joonhong","family":"Park","sequence":"additional","affiliation":[{"name":"Department of Laboratory Medicine, Jeonbuk National University Medical School and Hospital, Jeonju 54907, Republic of Korea"},{"name":"Research Institute of Clinical Medicine, Jeonbuk National University-Biomedical Research Institute, Jeonbuk National University Hospital, Jeonju 54907, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,2,17]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"84","DOI":"10.5045\/br.2017.52.2.84","article-title":"Diagnostic approaches for inherited hemolytic anemia in the genetic era","volume":"52","author":"Kim","year":"2017","journal-title":"Blood Res."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"3939","DOI":"10.1182\/blood-2008-07-161166","article-title":"Red cell membrane: Past, present, and future","volume":"112","author":"Mohandas","year":"2008","journal-title":"Blood"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"579","DOI":"10.1016\/j.pcl.2018.02.005","article-title":"Red Blood Cell Enzyme Disorders","volume":"65","author":"Grace","year":"2018","journal-title":"Pediatr. Clin. N. Am."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"6","DOI":"10.1093\/ajcp\/aqx047","article-title":"Molecular Diagnosis of Thalassemias and Hemoglobinopathies: An ACLPS Critical Review","volume":"148","author":"Sabath","year":"2017","journal-title":"Am. J. Clin. Pathol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"e279","DOI":"10.3346\/jkms.2020.35.e279","article-title":"Epidemiological Study of Hereditary Hemolytic Anemia in the Korean Pediatric Population during 1997-2016: A Nationwide Retrospective Cohort Study","volume":"35","author":"Shim","year":"2020","journal-title":"J. Korean Med. Sci."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"114","DOI":"10.1186\/s13023-019-1070-0","article-title":"Molecular diagnosis of hereditary spherocytosis by multi-gene target sequencing in Korea: Matching with osmotic fragility test and presence of spherocyte","volume":"14","author":"Choi","year":"2019","journal-title":"Orphanet. J. Rare Dis."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"281","DOI":"10.1080\/17474086.2021.1886919","article-title":"Screening tools for hereditary hemolytic anemia: New concepts and strategies","volume":"14","author":"Fermo","year":"2021","journal-title":"Expert Rev. Hematol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"672","DOI":"10.1002\/ajh.25058","article-title":"Multi-gene panel testing improves diagnosis and management of patients with hereditary anemias","volume":"93","author":"Russo","year":"2018","journal-title":"Am. J. Hematol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"684569","DOI":"10.3389\/fphys.2021.684569","article-title":"Targeted Next Generation Sequencing and Diagnosis of Congenital Hemolytic Anemias: A Three Years Experience Monocentric Study","volume":"12","author":"Fermo","year":"2021","journal-title":"Front. Physiol."},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Li, X., Buckton, A.J., Wilkinson, S.L., John, S., Walsh, R., Novotny, T., Valaskova, I., Gupta, M., Game, L., and Barton, P.J. (2013). Towards clinical molecular diagnosis of inherited cardiac conditions: A comparison of bench-top genome DNA sequencers. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0067744"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"163","DOI":"10.1007\/s12185-020-03010-9","article-title":"Characterization of hereditary red blood cell membranopathies using combined targeted next-generation sequencing and osmotic gradient ektacytometry","volume":"113","author":"Krishnevskaya","year":"2021","journal-title":"Int. J. Hematol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"405","DOI":"10.1038\/gim.2015.30","article-title":"Standards and guidelines for the interpretation of sequence variants: A joint consensus recommendation of the American College of Medical Genetics and Genomics and the Association for Molecular Pathology","volume":"17","author":"Richards","year":"2015","journal-title":"Genet. Med."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"321","DOI":"10.1080\/16078454.2020.1807227","article-title":"A novel EPB41 p.Trp704* mutation in a Korean patient with hereditary elliptocytosis: A case report","volume":"25","author":"Shin","year":"2020","journal-title":"Hematology"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"e9677","DOI":"10.1097\/MD.0000000000009677","article-title":"Targeted next-generation sequencing identifies a novel nonsense mutation in SPTB for hereditary spherocytosis: A case report of a Korean family","volume":"97","author":"Shin","year":"2018","journal-title":"Medicine"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"498","DOI":"10.1056\/NEJMra043442","article-title":"Diagnosis from the blood smear","volume":"353","author":"Bain","year":"2005","journal-title":"N. Engl. J. Med."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1111\/ijlh.12215","article-title":"Reporting and grading of abnormal red blood cell morphology","volume":"37","author":"Constantino","year":"2015","journal-title":"Int. J. Lab. Hematol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1408","DOI":"10.1111\/ijlh.13681","article-title":"Evaluation of Scopio Labs X100 Full Field PBS: The first high-resolution full field viewing of peripheral blood specimens combined with artificial intelligence-based morphological analysis","volume":"43","author":"Katz","year":"2021","journal-title":"Int. J. Lab. Hematol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"579","DOI":"10.1016\/j.jmoldx.2020.01.007","article-title":"Next-Generation Sequencing-Based Diagnosis of Unexplained Inherited Hemolytic Anemias Reveals Wide Genetic and Phenotypic Heterogeneity","volume":"22","author":"Jamwal","year":"2020","journal-title":"J. Mol. Diagn."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1284","DOI":"10.3324\/haematol.2016.142463","article-title":"New insights on hereditary erythrocyte membrane defects","volume":"101","author":"Andolfo","year":"2016","journal-title":"Haematologica"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"806","DOI":"10.1111\/bjh.14131","article-title":"Clinical utility of next-generation sequencing in the diagnosis of hereditary haemolytic anaemias","volume":"174","author":"Agarwal","year":"2016","journal-title":"Br. J. Haematol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"318","DOI":"10.1111\/bjh.14221","article-title":"A novel 33-Gene targeted resequencing panel provides accurate, clinical-grade diagnosis and improves patient management for rare inherited anaemias","volume":"175","author":"Roy","year":"2016","journal-title":"Br. J. Haematol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"388","DOI":"10.1111\/ijlh.12223","article-title":"Rare hereditary red blood cell enzymopathies associated with hemolytic anemia\u2014Pathophysiology, clinical aspects, and laboratory diagnosis","volume":"36","author":"Koralkova","year":"2014","journal-title":"Int. J. Lab. Hematol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"991","DOI":"10.1006\/bbrc.1995.1432","article-title":"Compound heterozygous mutations affecting both hepatic and erythrocyte isozymes of pyruvate kinase","volume":"208","author":"Uenaka","year":"1995","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"3585","DOI":"10.1182\/blood.V95.11.3585","article-title":"Estimating the prevalence of pyruvate kinase deficiency from the gene frequency in the general white population","volume":"95","author":"Beutler","year":"2000","journal-title":"Blood"},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Vives Corrons, J.L., Krishnevskaya, E., Montllor, L., Leguizamon, V., and Garcia Bernal, M. (2022). Concomitant Hereditary Spherocytosis and Pyruvate Kinase Deficiency in a Spanish Family with Chronic Hemolytic Anemia: Contribution of Laser Ektacytometry to Clinical Diagnosis. Cells, 11.","DOI":"10.3390\/cells11071133"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"E35","DOI":"10.1002\/ajh.23899","article-title":"Partial pyruvate kinase deficiency aggravates the phenotypic expression of band 3 deficiency in a family with hereditary spherocytosis","volume":"90","author":"Veldthuis","year":"2015","journal-title":"Am. J. Hematol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"386","DOI":"10.1111\/bjh.16084","article-title":"Hereditary spherocytosis is associated with decreased pyruvate kinase activity due to impaired structural integrity of the red blood cell membrane","volume":"187","author":"Andres","year":"2019","journal-title":"Br. J. Haematol."},{"key":"ref_28","first-page":"227","article-title":"Co-existence of hereditary spherocytosis and a new red cell pyruvate kinase variant: PK mallorca","volume":"85","author":"Zarza","year":"2000","journal-title":"Haematologica"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1645","DOI":"10.3346\/jkms.2013.28.11.1645","article-title":"Clinical characteristics of pediatric thalassemia in Korea: A single institute experience","volume":"28","author":"Hong","year":"2013","journal-title":"J. Korean Med. Sci."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"211","DOI":"10.5045\/br.2013.48.3.211","article-title":"Hereditary hemolytic anemia in Korea from 2007 to 2011: A study by the Korean Hereditary Hemolytic Anemia Working Party of the Korean Society of Hematology","volume":"48","author":"Park","year":"2013","journal-title":"Blood Res."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"133","DOI":"10.3343\/alm.2019.39.2.133","article-title":"Increased Prevalence of Thalassemia in Young People in Korea: Impact of Increasing Immigration","volume":"39","author":"Lee","year":"2019","journal-title":"Ann. Lab. Med."}],"container-title":["Diagnostics"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2075-4418\/13\/4\/770\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T18:39:48Z","timestamp":1760121588000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2075-4418\/13\/4\/770"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,2,17]]},"references-count":31,"journal-issue":{"issue":"4","published-online":{"date-parts":[[2023,2]]}},"alternative-id":["diagnostics13040770"],"URL":"https:\/\/doi.org\/10.3390\/diagnostics13040770","relation":{},"ISSN":["2075-4418"],"issn-type":[{"value":"2075-4418","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,2,17]]}}}