{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T04:38:44Z","timestamp":1787027924821,"version":"build-2736575974"},"reference-count":44,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2023,2,27]],"date-time":"2023-02-27T00:00:00Z","timestamp":1677456000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Universiti Sains Malaysia Research University","award":["1001\/PPSG\/8012366"],"award-info":[{"award-number":["1001\/PPSG\/8012366"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Diagnostics"],"abstract":"<jats:p>(1) Background: Alpha (\u03b1)-thalassaemia is a genetic disorder that affects 5% of the world population. Deletional or nondeletional mutations of one or both HBA1 and HBA2 on chromosome 16 will result in reduced production of \u03b1-globin chains, a component of haemoglobin (Hb) that is required for the formation of red blood cells (RBCs). This study aimed to determine the prevalence, haematological and molecular characterisations of \u03b1-thalassaemia. (2) Method: The parameters were based on full blood count, high-performance liquid chromatography and capillary electrophoresis. The molecular analysis involved gap-polymerase chain reaction (PCR), multiplex amplification refractory mutation system-PCR, multiplex ligation-dependent probe amplification and Sanger sequencing. (3) Results: With a total cohort of 131 patients, the prevalence of \u03b1-thalassaemia was 48.9%, leaving the remaining 51.1% with potentially undetected \u03b1 gene mutations. The following genotypes were detected: -\u03b13.7\/\u03b1\u03b1 (15.4%), -\u03b14.2\/\u03b1\u03b1 (3.7%), --SEA\/\u03b1\u03b1 (7.4%), \u03b1CS\u03b1\/\u03b1\u03b1 (10.3%), \u03b1Adana\u03b1\/\u03b1\u03b1 (0.7%), \u03b1Quong Sze\u03b1\/\u03b1\u03b1 (1.5%), -\u03b13.7\/-\u03b13.7 (0.7%), \u03b1CS\u03b1\/\u03b1CS\u03b1 (0.7%), -\u03b14.2\/\u03b1CS\u03b1 (0.7%), \u2013SEA\/\u03b1CS\u03b1 (1.5%), \u2013SEA\/\u03b1Quong Sze\u03b1 (0.7%), -\u03b13.7\/\u03b1Adana\u03b1 (0.7%), --SEA\/-\u03b13.7 (2.2%) and \u03b1CS\u03b1\/\u03b1Adana\u03b1 (0.7%). Indicators such as Hb (p = 0.022), mean corpuscular volume (p = 0.009), mean corpuscular haemoglobin (p = 0.017), RBC (p = 0.038) and haematocrit (p = 0.058) showed significant changes among patients with deletional mutations, but not between patients with nondeletional mutations. (4) Conclusions: A wide range of haematological parameters was observed among patients, including those with the same genotype. Thus, a combination of molecular technologies and haematological parameters is necessary for the accurate detection of \u03b1-globin chain mutations.<\/jats:p>","DOI":"10.3390\/diagnostics13050894","type":"journal-article","created":{"date-parts":[[2023,2,27]],"date-time":"2023-02-27T04:13:16Z","timestamp":1677471196000},"page":"894","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Gene Mutation Spectrum among Alpha-Thalassaemia Patients in Northeast Peninsular Malaysia"],"prefix":"10.3390","volume":"13","author":[{"given":"Divashini","family":"Vijian","sequence":"first","affiliation":[{"name":"School of Dental Sciences, Universiti Sains Malaysia, Kubang Kerian 16150, Kelantan, Malaysia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3878-208X","authenticated-orcid":false,"given":"Wan Suriana","family":"Wan Ab Rahman","sequence":"additional","affiliation":[{"name":"School of Dental Sciences, Universiti Sains Malaysia, Kubang Kerian 16150, Kelantan, Malaysia"},{"name":"Hospital Universiti Sains Malaysia, Kubang Kerian 16150, Kelantan, Malaysia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5015-9771","authenticated-orcid":false,"given":"Kannan Thirumulu","family":"Ponnuraj","sequence":"additional","affiliation":[{"name":"School of Dental Sciences, Universiti Sains Malaysia, Kubang Kerian 16150, Kelantan, Malaysia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zefarina","family":"Zulkafli","sequence":"additional","affiliation":[{"name":"Hospital Universiti Sains Malaysia, Kubang Kerian 16150, Kelantan, Malaysia"},{"name":"Department of Hematology, School of Medical Sciences, Universiti Sains Malaysia, Kubang Kerian 16150, Kelantan, Malaysia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rosnah","family":"Bahar","sequence":"additional","affiliation":[{"name":"Hospital Universiti Sains Malaysia, Kubang Kerian 16150, Kelantan, Malaysia"},{"name":"Department of Hematology, School of Medical Sciences, Universiti Sains Malaysia, Kubang Kerian 16150, Kelantan, Malaysia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4202-2438","authenticated-orcid":false,"given":"Norafiza","family":"Yasin","sequence":"additional","affiliation":[{"name":"Haematology Unit, Cancer Research Centre, Institute for Medical Research, Shah Alam 40170, Selangor, Malaysia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9886-504X","authenticated-orcid":false,"given":"Syahzuwan","family":"Hassan","sequence":"additional","affiliation":[{"name":"Haematology Unit, Cancer Research Centre, Institute for Medical Research, Shah Alam 40170, Selangor, Malaysia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ezalia","family":"Esa","sequence":"additional","affiliation":[{"name":"Haematology Unit, Cancer Research Centre, Institute for Medical Research, Shah Alam 40170, Selangor, Malaysia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,2,27]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Bain, B.J. (2020). Haemoglobinopathy Diagnosis, John Wiley & Sons.","DOI":"10.1002\/9781119579977"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"e2018054","DOI":"10.4084\/mjhid.2018.054","article-title":"Prevalence of thalassemia among newborns: A re-visited after 20 years of a prevention and control program in northeast Thailand","volume":"10","author":"Chaibunruang","year":"2018","journal-title":"Mediterr. J. Hematol. Infect. Dis."},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Goh, L.P.W., Chong, E.T.J., and Lee, P.-C. (2020). Prevalence of alpha (\u03b1)-thalassemia in Southeast Asia (2010\u20132020): A meta-analysis involving 83,674 subjects. Int. J. Environ. Res. Public Health, 17.","DOI":"10.3390\/ijerph17207354"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"e246062","DOI":"10.1590\/1519-6984.246062","article-title":"Thalassemia, a human blood disorder","volume":"83","author":"Shafique","year":"2021","journal-title":"Braz. J. Biol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"166","DOI":"10.1186\/s13023-020-01429-1","article-title":"Non-deletional alpha thalassaemia: A review","volume":"15","author":"Kwaifa","year":"2020","journal-title":"Orphanet J. Rare Dis."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"344","DOI":"10.4274\/tjh.2014.0204","article-title":"Evaluation of alpha-thalassemia mutations in cases with hypochromic microcytic anemia: The Istanbul perspective","volume":"32","author":"Karakas","year":"2015","journal-title":"Turk. J. Hematol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"327","DOI":"10.1089\/109065701753617471","article-title":"Detection of severe nondeletional \u03b1-thalassemia mutations using a single-tube multiplex ARMS assay","volume":"5","author":"Eng","year":"2001","journal-title":"Genet. Test."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"184","DOI":"10.3109\/03630261003670259","article-title":"A rapid single-tube multiplex polymerase chain reaction assay for the seven most prevalent \u03b1-thalassemia deletions and \u03b1\u03b1\u03b1anti 3.7 \u03b1-globin gene triplication","volume":"34","author":"Groeneger","year":"2010","journal-title":"Hemoglobin"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"e037974","DOI":"10.1136\/bmjopen-2020-037974","article-title":"Observational study on the current status of thalassaemia in Malaysia: A report from the Malaysian Thalassaemia Registry","volume":"10","author":"Ibrahim","year":"2020","journal-title":"BMJ Open"},{"key":"ref_10","first-page":"12","article-title":"Optimal mean corpuscular haemoglobin (MCH) cut-off value for differentiating alpha plus and alpha zero thalassaemia in thalassaemia screening","volume":"22","author":"Idris","year":"2020","journal-title":"Age (Mean \u00b1 SD)"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"18599","DOI":"10.3390\/ijms140918599","article-title":"Distribution of alpha thalassaemia gene variants in diverse ethnic populations in Malaysia: Data from the Institute for Medical Research","volume":"14","author":"Ahmad","year":"2013","journal-title":"Int. J. Mol. Sci."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Teh, L.K., Lim, L.F., Teh, Y.L., George, E., Lee, T., and Lim, L.N. (2020). Interaction of hematological analysis and \u03b1-globin genotypes among eligible blood donors. IIUM Med. J. Malays., 19.","DOI":"10.31436\/imjm.v19i3.1659"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"474","DOI":"10.1007\/s12185-019-02694-y","article-title":"Molecular characteristics of thalassemia and hemoglobin variants in prenatal diagnosis program in northern Thailand","volume":"110","author":"Mankhemthong","year":"2019","journal-title":"Int. J. Hematol."},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Pata, S., Laopajon, W., Pongpaiboon, M., Thongkum, W., Polpong, N., Munkongdee, T., Paiboonsukwong, K., Fucharoen, S., Tayapiwatana, C., and Kasinrerk, W. (2019). Impact of the detection of \u03b6-globin chains and hemoglobin Bart\u2019s using immunochromatographic strip tests for \u03b10-thalassemia (--SEA) differential diagnosis. PLoS ONE, 14.","DOI":"10.1371\/journal.pone.0223996"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"65","DOI":"10.1159\/000438739","article-title":"Molecular heterogeneity of thalassemia among pregnant Laotian women","volume":"135","author":"Wongprachum","year":"2016","journal-title":"Acta Haematol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"163","DOI":"10.3109\/03630269.2016.1158723","article-title":"Molecular epidemiology of hemoglobinopathies in Cambodia","volume":"40","author":"Munkongdee","year":"2016","journal-title":"Hemoglobin"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"161","DOI":"10.1016\/j.bcmd.2013.12.002","article-title":"Hemoglobin constant spring is markedly high in women of an ethnic minority group in Vietnam: A community-based survey and hematologic features","volume":"52","author":"Sanchaisuriya","year":"2014","journal-title":"Blood Cells Mol. Dis."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"359","DOI":"10.1111\/j.1365-2141.2010.08237.x","article-title":"Large scale screening for haemoglobin disorders in southern Vietnam: Implications for avoidance and management","volume":"150","author":"Hien","year":"2010","journal-title":"Br. J. Haematol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"557","DOI":"10.1080\/00365513.2021.1972332","article-title":"Direct PCR assays without DNA extraction for rapid detection of hemoglobin Constant Spring and Pakse\u2019 genes: Application for carrier screening and prenatal diagnosis","volume":"81","author":"Wichian","year":"2021","journal-title":"Scand. J. Clin. Lab. Investig."},{"key":"ref_20","first-page":"1","article-title":"Prevalence and disease burden of common alpha thalassemia deletions in Malaysian blood donors: A multi ethnic population","volume":"2","author":"Aliza","year":"2012","journal-title":"Int. J. Sci. Res."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"528","DOI":"10.1080\/00365513.2020.1795921","article-title":"Molecular spectrum of Hb H disease and characterization of rare deletional \u03b1-thalassemia found in Thailand","volume":"80","author":"Jomoui","year":"2020","journal-title":"Scand. J. Clin. Lab. Investig."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"271070","DOI":"10.1155\/2014\/271070","article-title":"Clinical features and molecular analysis of Hb H disease in Taiwan","volume":"2014","author":"Chao","year":"2014","journal-title":"BioMed Res. Int."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"43","DOI":"10.1016\/j.bcmd.2017.09.004","article-title":"Molecular basis of \u03b1-thalassemia","volume":"70","author":"Farashi","year":"2018","journal-title":"Blood Cells Mol. Dis."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"7329","DOI":"10.2147\/IJGM.S338419","article-title":"Screening of some indicators for alpha-thalassemia in Fujian province of Southern China","volume":"14","author":"Zheng","year":"2021","journal-title":"Int. J. Gen. Med."},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Osman, H.A., Hamid, M.M.A., Ahmad, R.B., Saleem, M., and Abdallah, S.A. (2020). Prevalence of 3.7 and 4.2 deletions in Sudanese patients with red cells hypochromia and microcytosis. BMC Res. Notes, 13.","DOI":"10.1186\/s13104-020-4933-5"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"32","DOI":"10.1111\/ijlh.12527","article-title":"Laboratory diagnosis of thalassemia","volume":"38","author":"Brancaleoni","year":"2016","journal-title":"Int. J. Lab. Hematol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"223","DOI":"10.1620\/tjem.223.223","article-title":"Elevated hemoglobin A2 as a marker for \u03b2-thalassemia trait in pregnant women","volume":"223","author":"Ou","year":"2011","journal-title":"Tohoku J. Exp. Med."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"102523","DOI":"10.1016\/j.bcmd.2020.102523","article-title":"MicroRNA expression patterns in HbE\/\u03b2-thalassemia patients: The passwords to unlock fetal hemoglobin expression in \u03b2-hemoglobinopathies","volume":"87","author":"Das","year":"2021","journal-title":"Blood Cells Mol. Dis."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"3844","DOI":"10.1038\/s41598-021-82297-y","article-title":"Comparison of cord blood hematological parameters among normal, \u03b1-thalassemia, and \u03b2-thalassemia fetuses between 17 and 38 weeks of gestation","volume":"11","author":"Zhan","year":"2021","journal-title":"Sci. Rep."},{"key":"ref_30","first-page":"339","article-title":"Alpha and beta thalassemia","volume":"80","author":"Campbell","year":"2009","journal-title":"Am. Fam. Physician"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"61","DOI":"10.1186\/s43042-022-00279-9","article-title":"Problem of borderline hemoglobin A2 levels in an Iranian population with a high prevalence of \u03b1-and \u03b2-thalassemia carriers","volume":"23","author":"Moradi","year":"2022","journal-title":"Egypt. J. Med. Hum. Genet."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"208","DOI":"10.1097\/MPH.0000000000001597","article-title":"Erythrocyte indices as differential diagnostic biomarkers of iron deficiency anemia and thalassemia","volume":"42","author":"Ozdemir","year":"2020","journal-title":"J. Pediatr. Hematol. Oncol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"S94","DOI":"10.1097\/01.PAT.0000443666.03931.9e","article-title":"Clinical and haematological features of non-deletional alpha thalassaemia mutations in Singapore","volume":"46","author":"Lam","year":"2014","journal-title":"Pathology"},{"key":"ref_34","first-page":"S124","article-title":"Prevalence and hematological parameters of thalassemia in Tha Kradarn subdistrict Chachoengsao Province, Thailand","volume":"95","author":"Areekul","year":"2012","journal-title":"J. Med. Assoc. Thail."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"175","DOI":"10.3109\/03630261003680191","article-title":"Detection of Hb Constant Spring by a capillary electrophoresis method","volume":"34","author":"Liao","year":"2010","journal-title":"Hemoglobin"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"254","DOI":"10.1007\/s12288-015-0532-3","article-title":"Hemoglobin constant spring (Hb CS) missed by HPLC in an Hb E trait pregnancy resulting in Hb H-CS disease in a Thai girl: Utility of capillary electrophoresis","volume":"32","author":"Pornprasert","year":"2016","journal-title":"Indian J. Hematol. Blood Transfus."},{"key":"ref_37","first-page":"21","article-title":"Molecular characterisation of haemoglobin constant spring and haemoglobin quong sze with a combine-amplification refractory mutation system","volume":"16","author":"Wee","year":"2009","journal-title":"Malays. J. Med. Sci."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"e27220","DOI":"10.1002\/pbc.27220","article-title":"Hb Adana (HBA2 or HBA1: C. 179G> A) and alpha thalassemia: Genotype\u2013phenotype correlation","volume":"65","author":"Singh","year":"2018","journal-title":"Pediatr. Blood Cancer"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"4856","DOI":"10.1038\/s41598-022-08986-4","article-title":"Genotype-phenotype correlation in patients with deletional and nondeletional mutations of Hb H disease in Southwest of Iran","volume":"12","author":"Hamid","year":"2022","journal-title":"Sci. Rep."},{"key":"ref_40","doi-asserted-by":"crossref","unstructured":"Shamoon, R.P., Yassin, A.K., Polus, R.K., and Ali, M.D. (2020). Genotype-phenotype correlation of HbH disease in northern Iraq. BMC Med. Genet., 21.","DOI":"10.1186\/s12881-020-01141-8"},{"key":"ref_41","first-page":"207","article-title":"[alpha]-Thalassemia with Haemoglobin Adana mutation: Prenatal diagnosis","volume":"36","author":"Zainal","year":"2014","journal-title":"Malays. J. Pathol."},{"key":"ref_42","first-page":"666","article-title":"Hemoglobin H disease in the eastern region of Saudi Arabia","volume":"21","author":"Qadri","year":"2000","journal-title":"Saudi Med. J."},{"key":"ref_43","unstructured":"Rifai, N. (2017). Tietz Textbook of Clinical Chemistry and Molecular Diagnostics-E-Book, Elsevier Health Sciences."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"333","DOI":"10.15562\/bmj.v8i2.1411","article-title":"A rare case of Hb H disease caused by compound heterozygous for \u03b1 thalasemia and Hb Quong Sze in Chinese Indonesian proband: A case report","volume":"8","author":"Widyastiti","year":"2019","journal-title":"Bali Med. J."}],"container-title":["Diagnostics"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2075-4418\/13\/5\/894\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T18:43:30Z","timestamp":1760121810000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2075-4418\/13\/5\/894"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,2,27]]},"references-count":44,"journal-issue":{"issue":"5","published-online":{"date-parts":[[2023,3]]}},"alternative-id":["diagnostics13050894"],"URL":"https:\/\/doi.org\/10.3390\/diagnostics13050894","relation":{},"ISSN":["2075-4418"],"issn-type":[{"value":"2075-4418","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,2,27]]}}}