{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,28]],"date-time":"2026-05-28T01:02:51Z","timestamp":1779930171037,"version":"3.53.1"},"reference-count":39,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2024,5,18]],"date-time":"2024-05-18T00:00:00Z","timestamp":1715990400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Conselho Nacional de Desenvolvimento Cient\u00edfico e Tecnol\u00f3gico (CNPQ)","award":["001"],"award-info":[{"award-number":["001"]}]},{"name":"Conselho Nacional de Desenvolvimento Cient\u00edfico e Tecnol\u00f3gico (CNPQ)","award":["2021"],"award-info":[{"award-number":["2021"]}]},{"name":"Coordena\u00e7\u00e3o de Aperfei\u00e7oamento de Pessoal de N\u00edvel Superior\u2014Brazil (CAPES)","award":["001"],"award-info":[{"award-number":["001"]}]},{"name":"Coordena\u00e7\u00e3o de Aperfei\u00e7oamento de Pessoal de N\u00edvel Superior\u2014Brazil (CAPES)","award":["2021"],"award-info":[{"award-number":["2021"]}]},{"name":"Programa Inova Fiocruz (Gera\u00e7\u00e3o de Conhecimento\u20142021)","award":["001"],"award-info":[{"award-number":["001"]}]},{"name":"Programa Inova Fiocruz (Gera\u00e7\u00e3o de Conhecimento\u20142021)","award":["2021"],"award-info":[{"award-number":["2021"]}]},{"name":"Funda\u00e7\u00e3o de Amparo a Pesquisa do Estado do Rio de Janeiro\/FAPERJ (APQ1\u20142022)","award":["001"],"award-info":[{"award-number":["001"]}]},{"name":"Funda\u00e7\u00e3o de Amparo a Pesquisa do Estado do Rio de Janeiro\/FAPERJ (APQ1\u20142022)","award":["2021"],"award-info":[{"award-number":["2021"]}]},{"name":"Programa de Incentivo \u00e0 Pesquisa (PIP IV)","award":["001"],"award-info":[{"award-number":["001"]}]},{"name":"Programa de Incentivo \u00e0 Pesquisa (PIP IV)","award":["2021"],"award-info":[{"award-number":["2021"]}]},{"name":"Rede de Laborat\u00f3rios de Refer\u00eancia-FIOCRUZ","award":["001"],"award-info":[{"award-number":["001"]}]},{"name":"Rede de Laborat\u00f3rios de Refer\u00eancia-FIOCRUZ","award":["2021"],"award-info":[{"award-number":["2021"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Genes"],"abstract":"<jats:p>Background: Diagnosing imprinting defects in neonates and young children presents challenges, often necessitating molecular analysis for a conclusive diagnosis. The isolation of genetic material from oral swabs becomes crucial, especially in settings where blood sample collection is impractical or for vulnerable populations like newborns, who possess limited blood volumes and are often too fragile for invasive procedures. Oral swab samples emerge as an excellent source of DNA, effectively overcoming obstacles associated with rare diseases. Methods: In our study, we specifically addressed the determination of the quality and quantity of DNA extracted from oral swab samples using NaCl procedures. Results: We compared these results with extractions performed using a commercial kit. Subsequently, the obtained material underwent MS\u2013HRM analysis for loci associated with imprinting diseases such as Prader\u2013Willi and Angelman syndromes. Conclusions: Our study emphasizes the significance of oral swab samples as a reliable source for obtaining DNA for MS\u2013HRM analysis. NaCl extraction stands out as a practical and cost-effective method for genetic studies, contributing to a molecular diagnosis that proves particularly beneficial for patients facing delays in characterization, ultimately influencing their treatment.<\/jats:p>","DOI":"10.3390\/genes15050641","type":"journal-article","created":{"date-parts":[[2024,5,20]],"date-time":"2024-05-20T11:06:41Z","timestamp":1716203201000},"page":"641","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Establishing a Standardized DNA Extraction Method Using NaCl from Oral Mucosa Cells for Its Application in Imprinting Diseases Such as Prader\u2013Willi and Angelman Syndromes: A Preliminary Investigation"],"prefix":"10.3390","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-7259-669X","authenticated-orcid":false,"given":"Let\u00edcia Lopes Cabral Guimar\u00e3es","family":"da 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Med. Genet."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1867","DOI":"10.1093\/hmg\/8.10.1867","article-title":"Towards a molecular understanding of Prader-Willi and Angelman syndromes","volume":"8","author":"Mann","year":"1999","journal-title":"Hum. Mol. Genet."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1086\/302473","article-title":"Genetics of Angelman Syndrome","volume":"65","author":"Jiang","year":"1999","journal-title":"Am. J. Hum. Genet."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"165","DOI":"10.4269\/ajtmh.2012.11-0713","article-title":"An Evaluation of Dried Blood Spots and Oral Swabs as Alternative Specimens for the Diagnosis of Dengue and Screening for Past Dengue Virus Exposure","volume":"87","author":"Anders","year":"2012","journal-title":"Am. J. Trop. Med. Hyg."},{"key":"ref_5","first-page":"101","article-title":"Evaluation of HBs-Ag and anti-HBc levels in serum and saliva of patients with hepatitis B","volume":"34","author":"Gharavi","year":"2020","journal-title":"Med. J. Islam. Repub. Iran"},{"key":"ref_6","unstructured":"(2008). Measles and rubella laboratory network: 2007 meeting on use of alternative sampling techniques for surveillance. Wkly. Epidemiol. Rec., 83, 229\u2013232."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"201","DOI":"10.1016\/S0033-3506(01)00444-9","article-title":"A comparison of oral fluid collection devices for use in the surveillance of virus diseases in children","volume":"115","author":"Vyse","year":"2001","journal-title":"Public Health."},{"key":"ref_8","first-page":"789","article-title":"Exploring the Efficacy of Buccal Swabs in DNA Collection for Sexual Assault Cases","volume":"64","author":"Raposo","year":"2019","journal-title":"J. Forensic Sci."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1319","DOI":"10.1007\/s00414-021-02601-3","article-title":"Suitability of specific soft tissue swabs for the forensic identification of highly decomposed bodies","volume":"135","author":"Helm","year":"2021","journal-title":"Int J Legal Med."},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Sakurada, K., Watanabe, K., and Akutsu, T. (2020). Current Methods for Body Fluid Identification Related to Sexual Crime: Focusing on Saliva, Semen, and Vaginal Fluid. Diagnostics, 10.","DOI":"10.3390\/diagnostics10090693"},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Han, J., Zhuang, B., Zou, L., Wang, D., Jiang, L., Wei, Y.L., Zhao, L., Zhao, L., and Li, C. (2024). A developmental validation of the Quick TargSeq 1.0 integrated system for automated DNA genotyping in forensic science for reference samples. Electrophoresis., 9.","DOI":"10.1002\/elps.202300187"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"101713","DOI":"10.1016\/j.legalmed.2020.101713","article-title":"Evaluation of Rapid DNA system for buccal swab and disaster victim identification samples","volume":"46","author":"Kitayama","year":"2020","journal-title":"Leg. Med."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"11","DOI":"10.1186\/s12939-022-01809-y","article-title":"Challenges and recommendations to increasing the use of exome sequencing and whole genome sequencing for diagnosing rare diseases in Brazil: An expert perspective","volume":"22","author":"Oliveira","year":"2023","journal-title":"Int. J. Equity Health"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"84","DOI":"10.1186\/s13023-022-02254-4","article-title":"Epidemiology of rare diseases in Brazil: Protocol of the Brazilian Rare Diseases Network (RARAS-BRDN)","volume":"17","author":"Artifon","year":"2022","journal-title":"Orphanet J. Rare Dis."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"789","DOI":"10.2217\/epi.16.8","article-title":"Imprinted genes and imprinting control regions show predominant intermediate methylation in adult somatic tissues","volume":"8","author":"Pervjakova","year":"2016","journal-title":"Epigenomics"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"429","DOI":"10.1186\/s13023-021-02061-3","article-title":"The IDeaS initiative: Pilot study to assess the impact of rare diseases on patients and healthcare systems","volume":"16","author":"Tisdale","year":"2021","journal-title":"Orphanet J. Rare Dis."},{"key":"ref_17","first-page":"305","article-title":"Molecular testing for imprinting disorders","volume":"32","author":"Beygo","year":"2020","journal-title":"Med. Genet."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"76","DOI":"10.1002\/(SICI)1096-8628(19990507)84:1<76::AID-AJMG16>3.0.CO;2-F","article-title":"Two cases of maternal uniparental disomy 14 with a phenotype overlapping with the Prader-Willi phenotype","volume":"84","author":"Berends","year":"1999","journal-title":"Am. J. Med. Genet."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1405","DOI":"10.1038\/ng.2776","article-title":"Truncating mutations of MAGEL2 cause Prader-Willi phenotypes and autism","volume":"45","author":"Schaaf","year":"2013","journal-title":"Nat. Genet."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1249","DOI":"10.1007\/s40618-015-0312-9","article-title":"Prader-Willi syndrome: A review of clinical, genetic, and endocrine findings","volume":"38","author":"Angulo","year":"2015","journal-title":"J. Endocrinol. Investig."},{"key":"ref_21","unstructured":"(2024, April 12). Genotypic and Phenotypic Relationship between Prader-Willi and Prader-Willi-Like Syndromes\u2014Genetics and Molecular Research (GMR) 2023. Available online: https:\/\/geneticsmr.com\/2023\/08\/31\/genotypic-and-phenotypic-relationship-between-prader-willi-and-prader-willi-like-syndromes\/."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"46","DOI":"10.21037\/tp.2016.07.04","article-title":"The dilemma of diagnostic testing for Prader-Willi syndrome","volume":"6","author":"Smith","year":"2017","journal-title":"Transl. Pediatr."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"511","DOI":"10.1358\/mf.2001.23.9.662139","article-title":"DNA research in forensic dentistry","volume":"23","author":"Lijnen","year":"2001","journal-title":"Methods Find. Exp. Clin. Pharmacol."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Okoniewski, K.C., Wheeler, A.C., Lee, S., Boyea, B., Raspa, M., Taylor, J.L., and Bailey, D.B. (2019). Early Identification of Fragile X Syndrome through Expanded Newborn Screening. Brain Sci., 9.","DOI":"10.3390\/brainsci9010004"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"e637","DOI":"10.1002\/mgg3.637","article-title":"A rapid and accurate methylation-sensitive high-resolution melting analysis assay for the diagnosis of Prader Willi and Angelman patients","volume":"7","author":"Ferreira","year":"2019","journal-title":"Mol. Genet. Genom. Med."},{"key":"ref_26","unstructured":"Adam, M.P., Feldman, J., Mirzaa, G.M., Pagon, R.A., Wallace, S.E., Bean, L.J.H., and Gripp, K.W. (2023, November 02). Prader-Willi Syndrome. 1998 Oct 6, Available online: https:\/\/www.ncbi.nlm.nih.gov\/books\/NBK1330\/."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"978","DOI":"10.1590\/S0004-27302005000600019","article-title":"Padroniza\u00e7\u00e3o da t\u00e9cnica de extra\u00e7\u00e3o de DNA de c\u00e9lulas de mucosa oral com NaCl: Aplica\u00e7\u00e3o no estudo do gene PROP1","volume":"49","author":"Billerbeck","year":"2005","journal-title":"Arq. Bras. Endocrinol. Metab."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"13026","DOI":"10.1038\/s41598-020-69750-0","article-title":"A newborn screening pilot study using methylation-sensitive high resolution melting on dried blood spots to detect Prader-Willi and Angelman syndromes","volume":"10","author":"Ferreira","year":"2020","journal-title":"Sci. Rep."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"10","DOI":"10.1016\/j.ab.2012.10.006","article-title":"Simultaneously extracting DNA, RNA, and protein using kits: Is sample quantity or quality prejudiced?","volume":"433","author":"Mathieson","year":"2013","journal-title":"Anal. Biochem."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"785","DOI":"10.1016\/j.beem.2016.11.003","article-title":"Diagnosis and treatment of GH deficiency in Prader\u2013Willi syndrome","volume":"30","author":"Grugni","year":"2016","journal-title":"Best Pract. Res. Clin. Endocrinol. Metab."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"405","DOI":"10.1001\/jamaneurol.2022.0067","article-title":"Multicenter Consensus Approach to Evaluation of Neonatal Hypotonia in the Genomic Era: A Review","volume":"79","author":"Morton","year":"2022","journal-title":"JAMA Neurol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"586","DOI":"10.1111\/j.1469-8749.2011.03918.x","article-title":"Genetic evaluation of the pediatric patient with hypotonia: Perspective from a hypotonia specialty clinic and review of the literature","volume":"53","author":"Lisi","year":"2011","journal-title":"Dev. Med. Child Neurol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1291","DOI":"10.1016\/B978-0-444-59565-2.00001-0","article-title":"Diagnostic workup for neuromuscular diseases","volume":"113","author":"Fardeau","year":"2013","journal-title":"Handb. Clin. Neurol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"1233158","DOI":"10.1126\/science.1233158","article-title":"Lentiviral hematopoietic stem cell gene therapy benefits metachromatic leukodystrophy","volume":"341","author":"Biffi","year":"2013","journal-title":"Science"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"71","DOI":"10.1016\/j.spen.2019.01.004","article-title":"Therapeutic Aspects in Congenital Myopathies","volume":"29","author":"Jungbluth","year":"2019","journal-title":"Semin. Pediatr. Neurol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"100","DOI":"10.1186\/s13148-020-00886-6","article-title":"Current status of development of methylation biomarkers for in vitro diagnostic IVD applications","volume":"12","author":"Sokolowska","year":"2020","journal-title":"Clin. Epigenetics"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"685123","DOI":"10.3389\/fgene.2021.685123","article-title":"A Protocol for Preconceptional Screening of Consanguineous Couples Using Whole Exome Sequencing","volume":"12","author":"Heller","year":"2021","journal-title":"Front. Genet."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"6944","DOI":"10.1038\/s41598-018-25311-0","article-title":"Quantitation of the cellular content of saliva and buccal swab samples","volume":"8","author":"Theda","year":"2018","journal-title":"Sci. Rep."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"1081424","DOI":"10.3389\/fgene.2023.1081424","article-title":"Microbial contamination and composition of oral samples subjected to clinical whole genome sequencing","volume":"14","author":"Kumar","year":"2023","journal-title":"Front Genet."}],"container-title":["Genes"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2073-4425\/15\/5\/641\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T14:44:37Z","timestamp":1760107477000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2073-4425\/15\/5\/641"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,5,18]]},"references-count":39,"journal-issue":{"issue":"5","published-online":{"date-parts":[[2024,5]]}},"alternative-id":["genes15050641"],"URL":"https:\/\/doi.org\/10.3390\/genes15050641","relation":{},"ISSN":["2073-4425"],"issn-type":[{"value":"2073-4425","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,5,18]]}}}