{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,5,14]],"date-time":"2025-05-14T00:36:41Z","timestamp":1747183001609,"version":"3.40.5"},"reference-count":35,"publisher":"Wiley","issue":"3","license":[{"start":{"date-parts":[[2023,1,29]],"date-time":"2023-01-29T00:00:00Z","timestamp":1674950400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Annals of Human Genetics"],"published-print":{"date-parts":[[2023,5]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Introduction<\/jats:title><jats:p>The \u03b1<jats:sup>0<\/jats:sup>\u2010thalassemia 44.6\u00a0kb or Chiang Rai (\u2010\u2010<jats:sup>CR<\/jats:sup>) deletion has been reported in northern Thailand and is capable of causing hemoglobin (Hb) H disease and a lethal \u03b1\u2010thalassemia genotype, Hb Bart's hydrops fetalis, in this region. However, there are no current data regarding the frequency of \u2010\u2010<jats:sup>CR<\/jats:sup> nationwide due to a lack of effective diagnostic assay. Therefore, this study aimed to develop a reliable platform for simultaneous genotyping of\u00a0\u2010\u2010<jats:sup>CR<\/jats:sup> and two common \u03b1<jats:sup>0<\/jats:sup>\u2010thalassemias in Thailand (\u2010\u2010<jats:sup>SEA<\/jats:sup> and \u2010\u2010<jats:sup>THAI<\/jats:sup>) and investigate the frequency of\u00a0\u2010\u2010<jats:sup>CR<\/jats:sup> across Thailand.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>Multiplex gap\u2010PCR assay and five renewable plasmid DNA controls for\u00a0\u2010\u2010<jats:sup>CR<\/jats:sup>,\u00a0\u2010\u2010<jats:sup>SEA<\/jats:sup>, \u2010\u2010<jats:sup>THAI<\/jats:sup>, \u03b12\u2010globin (<jats:italic>HBA2<\/jats:italic>), and \u03b2\u2010actin (<jats:italic>ACTB<\/jats:italic>) were newly developed and validated with reference methods. The developed assay was further tested on 1046 unrelated individuals with a reduced mean corpuscular volume (MCV) of less than 75 fl for investigating genotypic and allelic spectrum of \u2010\u2010<jats:sup>CR<\/jats:sup>.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Our developed assay showed 100% concordance with reference methods. The results were valid and reproducible throughout hundreds of reactions. Comparison of the genotypic and allelic spectra revealed that heterozygous \u2010\u2010<jats:sup>SEA<\/jats:sup> (\u2010\u2010<jats:sup>SEA<\/jats:sup>\/\u03b1\u03b1) and \u2010\u2010<jats:sup>SEA<\/jats:sup> alleles were dominant with the frequency of 22.85% (239\/1046) and 13.34% (279\/2092), respectively. Of these, \u2010\u2010<jats:sup>THAI<\/jats:sup> and \u2010\u2010<jats:sup>CR<\/jats:sup> were relatively rare in this population and comparable to each other with the allelic frequency of 0.14% (3\/2092).<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>This study successfully established a reliable molecular diagnostic platform for genotyping of \u2010\u2010<jats:sup>CR<\/jats:sup>, \u2010\u2010<jats:sup>SEA<\/jats:sup>, and \u2010\u2010<jats:sup>THAI<\/jats:sup> in a single reaction. Additionally, we demonstrated the frequency of \u2010\u2010<jats:sup>CR<\/jats:sup> in Thailand for the first time and provided knowledge basis for the planning of severe \u03b1\u2010thalassemia prevention and control programs in Thailand, where thalassemia is endemic.<\/jats:p><\/jats:sec>","DOI":"10.1111\/ahg.12496","type":"journal-article","created":{"date-parts":[[2023,1,29]],"date-time":"2023-01-29T10:48:20Z","timestamp":1674989300000},"page":"137-145","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Development of molecular diagnostic platform for \u03b1<sup>0<\/sup>\u2010thalassemia 44.6\u00a0kb (Chiang Rai, \u2010\u2010<sup>CR<\/sup>) deletion in individuals with microcytic red blood cells across Thailand"],"prefix":"10.1111","volume":"87","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-4597-6272","authenticated-orcid":false,"given":"Pinyaphat","family":"Khamphikham","sequence":"first","affiliation":[{"name":"Division of Clinical Microscopy, Department of Medical Technology, Faculty of Associated Medical Sciences Chiang Mai University  Chiang Mai Thailand"},{"name":"Hematology and Health Technology Research Center, Department of Medical Technology, Faculty of Associated Medical Sciences Chiang Mai University  Chiang Mai Thailand"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Oravee","family":"Hanmanoviriya","sequence":"additional","affiliation":[{"name":"Division of Clinical Microscopy, Department of Medical Technology, Faculty of Associated Medical Sciences Chiang Mai University  Chiang Mai Thailand"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Somsakul Pop","family":"Wongpalee","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Faculty of Medicine Chiang Mai University  Chiang Mai Thailand"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Thongperm","family":"Munkongdee","sequence":"additional","affiliation":[{"name":"Thalassemia Research Center, Institute of Molecular Biosciences Mahidol University  Nakhon Pathom Thailand"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kittiphong","family":"Paiboonsukwong","sequence":"additional","affiliation":[{"name":"Thalassemia Research Center, Institute of Molecular Biosciences Mahidol University  Nakhon Pathom Thailand"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yupin","family":"Jopang","sequence":"additional","affiliation":[{"name":"Regional Health Promotion Center 9 Nakhon Ratchasima, Department of Health Ministry of Public Health  Nakhon Ratchasima Thailand"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Chaowanee","family":"Wangchauy","sequence":"additional","affiliation":[{"name":"Hematology Unit, Department of Pathology, Faculty of Medicine Prince of Songkla University  Songkhla Thailand"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Charan","family":"Sancharernsook","sequence":"additional","affiliation":[{"name":"Department of Medical Technology Sunpasitthiprasong Hospital  Ubon Ratchathani Thailand"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nathawat","family":"Jinorose","sequence":"additional","affiliation":[{"name":"Department of Medical Technology Phayao Hospital  Phayao Thailand"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sakorn","family":"Pornprasert","sequence":"additional","affiliation":[{"name":"Division of Clinical Microscopy, Department of Medical Technology, Faculty of Associated Medical Sciences Chiang Mai University  Chiang Mai Thailand"},{"name":"Hematology and Health Technology Research Center, Department of Medical Technology, Faculty of Associated Medical Sciences Chiang Mai University  Chiang Mai Thailand"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2023,1,29]]},"reference":[{"key":"e_1_2_8_2_1","doi-asserted-by":"publisher","DOI":"10.1186\/s13059-017-1212-4"},{"key":"e_1_2_8_3_1","doi-asserted-by":"publisher","DOI":"10.1182\/hematology.2020000158"},{"key":"e_1_2_8_4_1","doi-asserted-by":"publisher","DOI":"10.1080\/03014460500075084"},{"key":"e_1_2_8_5_1","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMra1215361"},{"key":"e_1_2_8_6_1","doi-asserted-by":"publisher","DOI":"10.1111\/ijlh.12082"},{"key":"e_1_2_8_7_1","doi-asserted-by":"publisher","DOI":"10.1097\/GIM.0b013e3181fcb468"},{"key":"e_1_2_8_8_1","doi-asserted-by":"publisher","DOI":"10.1093\/cdn\/nzaa053_035"},{"key":"e_1_2_8_9_1","doi-asserted-by":"publisher","DOI":"10.3390\/ijerph17207354"},{"volume-title":"Laboratory diagnosis of thalassemia and hemoglobinopathy","year":"2015","author":"Guidelines Review Committee. 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