{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,9,25]],"date-time":"2025-09-25T18:10:45Z","timestamp":1758823845766,"version":"3.37.3"},"reference-count":38,"publisher":"Public Library of Science (PLoS)","issue":"4","license":[{"start":{"date-parts":[[2024,4,10]],"date-time":"2024-04-10T00:00:00Z","timestamp":1712707200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100015747","name":"University Grants Commission of Bangladesh","doi-asserted-by":"publisher","award":["CP-4029"],"award-info":[{"award-number":["CP-4029"]}],"id":[{"id":"10.13039\/100015747","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS ONE"],"abstract":"<jats:p><jats:italic>TPO<\/jats:italic> (Thyroid Peroxidase) is known to be one of the major genes involved in congenital hypothyroid patients with thyroid dyshormonogenesis. The present study aims to validate high-resolution melting (HRM) curve analysis as a substitute method for Sanger sequencing, focusing on the frequently observed non-synonymous mutations c.1117G&gt;T, c.1193G&gt;C, and c.2173A&gt;C in the <jats:italic>TPO<\/jats:italic> gene in patients from Bangladesh. We enrolled 36 confirmed cases of congenital hypothyroid patients with dyshormonogenesis to establish the HRM method. Blood specimens were collected, and DNA was extracted followed by PCR and Sanger sequencing. Among the 36 specimens, 20 were pre-sequenced, and variants were characterized through Sanger sequencing. Following pre-sequencing, the 20 pre-sequenced specimens underwent real-time PCR-HRM curve analysis to determine the proper HRM condition for separating the three variations from the wild-type state into heterozygous and homozygous states. Furthermore, 16 unknown specimens were subjected to HRM analysis to validate the method. This method demonstrated a sensitivity and specificity of 100 percent in accurately discerning wild-type alleles from both homozygous and heterozygous states of c.1117G&gt;T (23\/36; 63.8%), c.1193G&gt;C (30\/36; 83.3%), and c.2173A&gt;C (23\/36; 63.8%) variants frequently encountered among 36 Bangladeshi patients. The HRM data was found to be similar to the sequencing result, thus confirming the validity of the HRM approach for <jats:italic>TPO<\/jats:italic> gene variant detection. In conclusion, HRM-based molecular technique targeting variants c.1117G&gt;T, c.1193G&gt;C, and c.2173A&gt;C could be used as a high throughput, rapid, reliable, and cost-effective screening approach for the detection of all common mutations in <jats:italic>TPO<\/jats:italic> gene in Bangladeshi patients with dyshormonogenesis.<\/jats:p>","DOI":"10.1371\/journal.pone.0293570","type":"journal-article","created":{"date-parts":[[2024,4,10]],"date-time":"2024-04-10T17:30:42Z","timestamp":1712770242000},"page":"e0293570","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":2,"title":["High-resolution melt curve analysis: An approach for variant detection in the TPO gene of congenital hypothyroid patients in Bangladesh"],"prefix":"10.1371","volume":"19","author":[{"given":"Mst. Noorjahan","family":"Begum","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rumana","family":"Mahtarin","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2822-245X","authenticated-orcid":true,"given":"Md Tarikul","family":"Islam","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nusrat Jahan","family":"Antora","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Suprovath Kumar","family":"Sarker","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nusrat","family":"Sultana","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1710-9865","authenticated-orcid":true,"given":"Abu A.","family":"Sajib","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7274-0855","authenticated-orcid":true,"given":"Abul B. M. M. K.","family":"Islam","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hurjahan","family":"Banu","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"M. A.","family":"Hasanat","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kohinoor Jahan","family":"Shyamaly","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Suraiya","family":"Begum","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tasnia Kawsar","family":"Konika","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shahinur","family":"Haque","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mizanul","family":"Hasan","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sadia","family":"Sultana","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3755-4763","authenticated-orcid":true,"given":"Taufiqur Rahman","family":"Bhuiyan","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kaiissar","family":"Mannoor","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Firdausi","family":"Qadri","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0974-7667","authenticated-orcid":true,"given":"Sharif","family":"Akhteruzzaman","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"340","published-online":{"date-parts":[[2024,4,10]]},"reference":[{"issue":"1","key":"pone.0293570.ref001","first-page":"33","article-title":"Congenital hypothyroidism: Orphanet J Rare Dis","volume":"14","author":"MV Rastogi","year":"2012","journal-title":"Journal of Continuing Education Topics & Issues"},{"issue":"4","key":"pone.0293570.ref002","doi-asserted-by":"crossref","DOI":"10.5812\/ijem.38256","article-title":"Permanent and transient congenital hypothyroidism in Hamadan West Province of Iran","volume":"14","author":"Z Razavi","year":"2016","journal-title":"International journal of endocrinology and metabolism"},{"issue":"2","key":"pone.0293570.ref003","doi-asserted-by":"crossref","first-page":"104","DOI":"10.1038\/nrendo.2011.160","article-title":"Detection and treatment of congenital hypothyroidism","volume":"8","author":"A Gr\u00fcters","year":"2012","journal-title":"Nature Reviews Endocrinology"},{"issue":"5","key":"pone.0293570.ref004","doi-asserted-by":"crossref","first-page":"559","DOI":"10.1515\/JPEM.2007.20.5.559","article-title":"How should we be treating children with congenital hypothyroidism?","volume":"20","author":"SH LaFranchi","year":"2007","journal-title":"Journal of Pediatric Endocrinology and Metabolism"},{"issue":"S 04","key":"pone.0293570.ref005","doi-asserted-by":"crossref","first-page":"19","DOI":"10.1055\/s-0029-1211926","article-title":"Epidemiology of congenital hypothyroidism","volume":"105","author":"M Klett","year":"1997","journal-title":"Experimental and Clinical Endocrinology & Diabetes"},{"issue":"3","key":"pone.0293570.ref006","doi-asserted-by":"crossref","first-page":"268","DOI":"10.1016\/j.ymgme.2007.03.012","article-title":"Increase in congenital hypothyroidism in New York State and in the United States","volume":"91","author":"KB Harris","year":"2007","journal-title":"Molecular genetics and metabolism"},{"issue":"8","key":"pone.0293570.ref007","doi-asserted-by":"crossref","first-page":"3158","DOI":"10.1210\/jc.2007-0527","article-title":"Random variability in congenital hypothyroidism from thyroid dysgenesis over 16 years in Quebec","volume":"92","author":"J Delado\u00eby","year":"2007","journal-title":"The Journal of Clinical Endocrinology & Metabolism"},{"issue":"Supplement 2","key":"pone.0293570.ref008","doi-asserted-by":"crossref","first-page":"S31","DOI":"10.1542\/peds.2009-1975C","article-title":"Prevalence of congenital hypothyroidism\u2014current trends and future directions: workshop summary","volume":"125","author":"RS Olney","year":"2010","journal-title":"Pediatrics"},{"key":"pone.0293570.ref009","first-page":"162","article-title":"Screening for congenital hypothyroidism-a new era in Bangladesh","volume":"34","author":"M Hasan","year":"2004","journal-title":"SOUTHEAST ASIAN JOURNAL OF TROPICAL MEDICINE AND PUBLIC HEALTH"},{"issue":"5","key":"pone.0293570.ref010","doi-asserted-by":"crossref","first-page":"379","DOI":"10.1136\/jmg.2004.024158","article-title":"Genetics of congenital hypothyroidism","volume":"42","author":"S Park","year":"2005","journal-title":"Journal of medical genetics"},{"key":"pone.0293570.ref011","volume":"2019","author":"M Begum","year":"2019","journal-title":"Mutation spectrum in TPO gene of Bangladeshi patients with thyroid dyshormonogenesis and analysis of the effects of different mutations on the structural features and functions of TPO protein through in silico approach"},{"issue":"9","key":"pone.0293570.ref012","first-page":"e0291386","volume":"18","author":"MN Begum","year":"2023","journal-title":"Investigation of the impact of nonsynonymous mutations on thyroid peroxidase dimer"},{"issue":"8","key":"pone.0293570.ref013","first-page":"e0282553","volume":"18","author":"MN Begum","year":"2023","journal-title":"Molecular investigation of TSHR gene in Bangladeshi congenital hypothyroid patients"},{"issue":"4","key":"pone.0293570.ref014","doi-asserted-by":"crossref","first-page":"421","DOI":"10.1097\/MOP.0b013e32834726a4","article-title":"Genetic causes of congenital hypothyroidism due to dyshormonogenesis","volume":"23","author":"H Grasberger","year":"2011","journal-title":"Current opinion in pediatrics"},{"issue":"5","key":"pone.0293570.ref015","doi-asserted-by":"crossref","first-page":"511","DOI":"10.1530\/EJE-07-0037","article-title":"High prevalence of thyroid peroxidase gene mutations in patients with thyroid dyshormonogenesis","volume":"156","author":"M Avbelj","year":"2007","journal-title":"European journal of endocrinology"},{"issue":"3","key":"pone.0293570.ref016","doi-asserted-by":"crossref","first-page":"259","DOI":"10.1002\/humu.9175","article-title":"Five novel inactivating mutations in the thyroid peroxidase gene responsible for congenital goiter and iodide organification defect","volume":"22","author":"CM Rivolta","year":"2003","journal-title":"Human mutation"},{"issue":"10","key":"pone.0293570.ref017","doi-asserted-by":"crossref","first-page":"3708","DOI":"10.1210\/jcem.85.10.6878","article-title":"Two decades of screening for congenital hypothyroidism in The Netherlands: TPO gene mutations in total iodide organification defects (an update)","volume":"85","author":"B Bakker","year":"2000","journal-title":"The Journal of Clinical Endocrinology & Metabolism"},{"issue":"2","key":"pone.0293570.ref018","doi-asserted-by":"crossref","first-page":"198","DOI":"10.1046\/j.1365-2265.2003.01823.x","article-title":"Partial iodide organification defect caused by a novel mutation of the thyroid peroxidase gene in three siblings","volume":"59","author":"T Kotani","year":"2003","journal-title":"Clinical endocrinology"},{"issue":"2","key":"pone.0293570.ref019","doi-asserted-by":"crossref","first-page":"193","DOI":"10.1530\/eje.1.01826","article-title":"Mutation screening of the thyroid peroxidase gene in a cohort of 55 Portuguese patients with congenital hypothyroidism","volume":"152","author":"C Rodrigues","year":"2005","journal-title":"European Journal of Endocrinology"},{"issue":"3","key":"pone.0293570.ref020","doi-asserted-by":"crossref","first-page":"627","DOI":"10.1677\/joe.0.1720627","article-title":"Mutation analysis of thyroid peroxidase gene in Chinese patients with total iodide organification defect: identification of five novel mutations","volume":"172","author":"J Wu","year":"2002","journal-title":"Journal of Endocrinology"},{"issue":"3","key":"pone.0293570.ref021","doi-asserted-by":"crossref","first-page":"289","DOI":"10.1507\/endocrj.EJ13-0237","article-title":"Genetic analysis of thyroid peroxidase (TPO) gene in patients whose hypothyroidism was found in adulthood in West Bengal, India","volume":"61","author":"N Balmiki","year":"2014","journal-title":"Endocrine journal"},{"issue":"11","key":"pone.0293570.ref022","doi-asserted-by":"crossref","first-page":"1291","DOI":"10.1515\/JPEM.2006.19.11.1291","article-title":"Newborn screening for congenital hypothyroidism","volume":"19","author":"A. B\u00fcy\u00fckgebiz","year":"2006","journal-title":"Journal of Pediatric Endocrinology and Metabolism"},{"issue":"3","key":"pone.0293570.ref023","doi-asserted-by":"crossref","first-page":"305","DOI":"10.2353\/jmoldx.2010.090104","article-title":"Rapid and reliable detection of glucose-6-phosphate dehydrogenase (G6PD) gene mutations in Han Chinese using high-resolution melting analysis","volume":"12","author":"JB Yan","year":"2010","journal-title":"The Journal of Molecular Diagnostics"},{"issue":"1","key":"pone.0293570.ref024","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1186\/s12863-017-0594-3","article-title":"High resolution melting curve analysis targeting the HBB gene mutational hot-spot offers a reliable screening approach for all common as well as most of the rare beta-globin gene mutations in Bangladesh","volume":"19","author":"MT Islam","year":"2018","journal-title":"BMC genetics"},{"issue":"1","key":"pone.0293570.ref025","doi-asserted-by":"crossref","first-page":"323","DOI":"10.1186\/1756-0500-4-323","article-title":"Genetic screening of Fabry patients with EcoTILLING and HRM technology","volume":"4","author":"C Bono","year":"2011","journal-title":"BMC research notes"},{"issue":"1","key":"pone.0293570.ref026","doi-asserted-by":"crossref","first-page":"57","DOI":"10.1186\/1471-2369-12-57","article-title":"High Resolution Melt analysis for mutation screening in PKD1 and PKD2","volume":"12","author":"S Bataille","year":"2011","journal-title":"BMC nephrology"},{"issue":"6","key":"pone.0293570.ref027","doi-asserted-by":"crossref","first-page":"711","DOI":"10.1007\/s40618-017-0795-7","article-title":"High-resolution melting analysis (HRM) for mutational screening of Dnajc17 gene in patients affected by thyroid dysgenesis","volume":"41","author":"IC Nettore","year":"2018","journal-title":"Journal of endocrinological investigation"},{"key":"pone.0293570.ref028","doi-asserted-by":"crossref","first-page":"548","DOI":"10.33314\/jnhrc.v17i4.1042","article-title":"The Declaration of Helsinki on medical research involving human subjects: a review of seventh revision (Medical Education)","volume":"17","author":"B Shrestha","year":"2020","journal-title":"Journal of Nepal Health Research Council"},{"key":"pone.0293570.ref029","article-title":"Prevention of intellectual disability through screening for congenital hypothyroidism: how much and at what level?","author":"SD Grosse","year":"2011","journal-title":"Archives of disease in childhood"},{"issue":"3","key":"pone.0293570.ref030","first-page":"1061","article-title":"Two different mutations in the thyroid peroxidase gene of a large inbred Amish kindred: power and limits of homozygosity mapping","volume":"84","author":"S Pannain","year":"1999","journal-title":"The Journal of Clinical Endocrinology & Metabolism"},{"issue":"9","key":"pone.0293570.ref031","doi-asserted-by":"crossref","first-page":"1085","DOI":"10.1089\/thy.2005.15.1085","article-title":"Total iodide organification defect: clinical and molecular characterization of an Italian family","volume":"15","author":"L Fugazzola","year":"2005","journal-title":"Thyroid"},{"issue":"1","key":"pone.0293570.ref032","doi-asserted-by":"crossref","first-page":"82","DOI":"10.1006\/abbi.1994.1474","article-title":"Evidence for a radical mechanism in peroxidase-catalyzed coupling. I. Steady-state experiments with various peroxidases","volume":"315","author":"A Taurog","year":"1994","journal-title":"Archives of biochemistry and biophysics"},{"issue":"4","key":"pone.0293570.ref033","doi-asserted-by":"crossref","first-page":"568","DOI":"10.1111\/j.1365-2265.2011.04249.x","article-title":"Congenital goitrous hypothyroidism: mutation analysis in the thyroid peroxidase gene","volume":"76","author":"FS Belforte","year":"2012","journal-title":"Clinical endocrinology"},{"issue":"1","key":"pone.0293570.ref034","doi-asserted-by":"crossref","first-page":"9","DOI":"10.1002\/humu.1380060104","article-title":"Identification of five novel inactivating mutations in the human thyroid peroxidase gene by denaturing gradient gel electrophoresis","volume":"6","author":"H Bikker","year":"1995","journal-title":"Human Mutation"},{"issue":"10","key":"pone.0293570.ref035","doi-asserted-by":"crossref","first-page":"4481","DOI":"10.1021\/bi00436a054","article-title":"Structure of the human thyroid peroxidase gene: comparison and relationship to the human myeloperoxidase gene","volume":"28","author":"S Kimura","year":"1989","journal-title":"Biochemistry"},{"key":"pone.0293570.ref036","doi-asserted-by":"crossref","DOI":"10.1155\/2014\/390121","article-title":"Functional analysis of thyroid peroxidase gene mutations detected in patients with thyroid dyshormonogenesis","volume":"2014","author":"S Guria","year":"2014","journal-title":"International journal of endocrinology"},{"issue":"1","key":"pone.0293570.ref037","doi-asserted-by":"crossref","first-page":"58","DOI":"10.1186\/s12863-018-0664-1","article-title":"High resolution melting curve analysis enables rapid and reliable detection of G6PD variants in heterozygous females","volume":"19","author":"MT Islam","year":"2018","journal-title":"BMC genetics"},{"issue":"11","key":"pone.0293570.ref038","doi-asserted-by":"crossref","first-page":"2316","DOI":"10.3390\/ijms18112316","article-title":"High Resolution Melting (HRM) for High-Throughput Genotyping\u2014Limitations and Caveats in Practical Case Studies","volume":"18","author":"M S\u0142omka","year":"2017","journal-title":"International journal of molecular sciences"}],"container-title":["PLOS ONE"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0293570","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,4,10]],"date-time":"2024-04-10T17:31:09Z","timestamp":1712770269000},"score":1,"resource":{"primary":{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0293570"}},"subtitle":[],"editor":[{"given":"Yevgenya","family":"Grinblat","sequence":"first","affiliation":[],"role":[{"role":"editor","vocabulary":"crossref"}]}],"short-title":[],"issued":{"date-parts":[[2024,4,10]]},"references-count":38,"journal-issue":{"issue":"4","published-online":{"date-parts":[[2024,4,10]]}},"URL":"https:\/\/doi.org\/10.1371\/journal.pone.0293570","relation":{},"ISSN":["1932-6203"],"issn-type":[{"type":"electronic","value":"1932-6203"}],"subject":[],"published":{"date-parts":[[2024,4,10]]}}}