{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,7]],"date-time":"2026-07-07T22:07:53Z","timestamp":1783462073825,"version":"3.55.0"},"reference-count":45,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2021,8,11]],"date-time":"2021-08-11T00:00:00Z","timestamp":1628640000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100006314","name":"Biogen Idec","doi-asserted-by":"publisher","award":["BRA-SMG-17-11235"],"award-info":[{"award-number":["BRA-SMG-17-11235"]}],"id":[{"id":"10.13039\/100006314","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001807","name":"Funda\u00e7\u00e3o de Amparo \u00e0 Pesquisa do Estado de S\u00e3o Paulo","doi-asserted-by":"publisher","award":["14\/50931-3"],"award-info":[{"award-number":["14\/50931-3"]}],"id":[{"id":"10.13039\/501100001807","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJNS"],"abstract":"<jats:p>Since the approval of modifying therapies for Spinal Muscular Atrophy (SMA), several protocols aiming to screen SMN1 homozygous deletion in a neonatal context have been published. However, no work has compared different methodologies along with detailed implementation costs for centers where the neonatal screening of SMA has not yet been implemented. Therefore, our work compared different qualitative real-time PCR approaches for SMA screening and the estimated costs of test implementation. Using Brazilian blood samples, the presence and absence (P\/A) and melt curve protocols were analyzed. MLPA was used as a confirmatory test. The costs were calculated for the simplex and multiplex tests plus equipment. The test workflow was based on the present experience and literature report. The accuracy of the P\/A protocol was 1 (95% CI 0.8677\u22121) using dried blood spots (DBS). The melt curve protocol also achieved 100% concordance. The consumable costs ranged from USD 1.68 to 4.42 and from USD 2.04 to 12.76 per reaction, for the simplex and multiplex tests, respectively. The equipment acquisition costs ranged from USD 44,817.07 to 467,253.10, with several factors influencing this value presented. Our work presents a framework for decision-making, with a project demonstration of the different assays that will be useful in dealing with the issues of cost and availability of reagents. Moreover, we present a literature review and discussion of important concerns regarding treatment policies. We take the first step towards a future SMA NBS pilot program where it is not yet a reality.<\/jats:p>","DOI":"10.3390\/ijns7030053","type":"journal-article","created":{"date-parts":[[2021,8,11]],"date-time":"2021-08-11T04:56:22Z","timestamp":1628657782000},"page":"53","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":14,"title":["Newborn Screening for 5q Spinal Muscular Atrophy: Comparisons between Real-Time PCR Methodologies and Cost Estimations for Future Implementation Programs"],"prefix":"10.3390","volume":"7","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-8845-5952","authenticated-orcid":false,"given":"Vanessa Luiza","family":"Romanelli Tavares","sequence":"first","affiliation":[{"name":"Human Genome and Stem Cell Research Center, Department of Genetics and Evolutionary Biology, Institute of Bioscience, University of S\u00e3o Paulo, S\u00e3o Paulo 05508-090, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Frederico","family":"Monfardini","sequence":"additional","affiliation":[{"name":"Human Genome and Stem Cell Research Center, Department of Genetics and Evolutionary Biology, Institute of Bioscience, University of S\u00e3o Paulo, S\u00e3o Paulo 05508-090, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Naila Cristina Vila\u00e7a","family":"Louren\u00e7o","sequence":"additional","affiliation":[{"name":"Human Genome and Stem Cell Research Center, Department of Genetics and Evolutionary Biology, Institute of Bioscience, University of S\u00e3o Paulo, S\u00e3o Paulo 05508-090, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Katia Maria","family":"da Rocha","sequence":"additional","affiliation":[{"name":"Human Genome and Stem Cell Research Center, Department of Genetics and Evolutionary Biology, Institute of Bioscience, University of S\u00e3o Paulo, S\u00e3o Paulo 05508-090, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Karina","family":"Weinmann","sequence":"additional","affiliation":[{"name":"Human Genome and Stem Cell Research Center, Department of Genetics and Evolutionary Biology, Institute of Bioscience, University of S\u00e3o Paulo, S\u00e3o Paulo 05508-090, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rita","family":"Pavanello","sequence":"additional","affiliation":[{"name":"Human Genome and Stem Cell Research Center, Department of Genetics and Evolutionary Biology, Institute of Bioscience, University of S\u00e3o Paulo, S\u00e3o Paulo 05508-090, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mayana","family":"Zatz","sequence":"additional","affiliation":[{"name":"Human Genome and Stem Cell Research Center, Department of Genetics and Evolutionary Biology, Institute of Bioscience, University of S\u00e3o Paulo, S\u00e3o Paulo 05508-090, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2021,8,11]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"865","DOI":"10.1111\/j.1651-2227.2008.01201.x","article-title":"A population-based study of genotypic and phenotypic variability in children with spinal muscular atrophy","volume":"98","author":"Arkblad","year":"2009","journal-title":"Acta Paediatr. Int. J. Paediatr."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"27","DOI":"10.1038\/ejhg.2011.134","article-title":"Pan-ethnic carrier screening and prenatal diagnosis for spinal muscular atrophy: Clinical laboratory analysis of >72,400 specimens","volume":"20","author":"Sugarman","year":"2012","journal-title":"Eur. J. Hum. Genet."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1186\/s13023-017-0671-8","article-title":"Prevalence, incidence and carrier frequency of 5q-linked spinal muscular atrophy-A literature review","volume":"12","author":"Verhaart","year":"2017","journal-title":"Orphanet J. Rare Dis."},{"key":"ref_4","first-page":"39","article-title":"Advances in Newborn Screening and Presymptomatic Diagnosis of Spinal Muscular Atrophy","volume":"10","year":"2020","journal-title":"Degener. Neurol. Neuromuscul. Dis."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"2120","DOI":"10.1016\/S0140-6736(08)60921-6","article-title":"Spinal muscular atrophy","volume":"371","author":"Lunn","year":"2008","journal-title":"Lancet"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"71","DOI":"10.1186\/1750-1172-6-71","article-title":"Spinal muscular atrophy","volume":"6","author":"Mercuri","year":"2011","journal-title":"Orphanet J. Rare Dis."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"197","DOI":"10.1016\/j.nmd.2017.11.004","article-title":"Diagnosis and management of spinal muscular atrophy: Part 2: Pulmonary and acute care; medications, supplements and immunizations; other organ systems; and ethics","volume":"28","author":"Finkel","year":"2018","journal-title":"Neuromuscul. Disord."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"228","DOI":"10.1002\/(SICI)1098-1004(200003)15:3<228::AID-HUMU3>3.0.CO;2-9","article-title":"An update of the mutation spectrum of the survival motor neuron gene (SMN1) in autosomal recessive spinal muscular atrophy (SMA)","volume":"15","author":"Wirth","year":"2000","journal-title":"Hum. Mutat."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"696","DOI":"10.1097\/MOP.0b013e32833f3046","article-title":"Spinal muscular atrophy: A time for screening","volume":"22","author":"Prior","year":"2010","journal-title":"Curr. Opin. Pediatr."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"339","DOI":"10.2174\/1389202919666180101154916","article-title":"Molecular Factors Involved in Spinal Muscular Atrophy Pathways as Possible Disease-modifying Candidates","volume":"19","author":"Maretina","year":"2018","journal-title":"Curr. Genom."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"307","DOI":"10.1080\/14656566.2019.1704732","article-title":"New treatments in spinal muscular atrophy: An overview of currently available data","volume":"21","author":"Ramdas","year":"2020","journal-title":"Expert Opin. Pharmacother."},{"key":"ref_12","first-page":"194","article-title":"Nusinersen: A novel antisense oligonucleotide for the treatment of spinal muscular atrophy","volume":"24","author":"Neil","year":"2019","journal-title":"J. Pediatr. Pharmacol. Ther."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1002\/prp2.447","article-title":"Risdiplam distributes and increases SMN protein in both the central nervous system and peripheral organs","volume":"6","author":"Poirier","year":"2018","journal-title":"Pharmacol. Res. Perspect."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1001","DOI":"10.1177\/1060028020914274","article-title":"Onasemnogene Abeparvovec-xioi: Gene Therapy for Spinal Muscular Atrophy","volume":"54","author":"Stevens","year":"2020","journal-title":"Ann. Pharmacother."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"704","DOI":"10.1002\/ana.20473","article-title":"Natural history of denervation in SMA: Relation to age, SMN2 copy number, and function","volume":"57","author":"Swoboda","year":"2005","journal-title":"Ann. Neurol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"6307","DOI":"10.1007\/s12035-017-0831-9","article-title":"Time Is Motor Neuron: Therapeutic Window and Its Correlation with Pathogenetic Mechanisms in Spinal Muscular Atrophy","volume":"55","author":"Govoni","year":"2018","journal-title":"Mol. Neurobiol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"124","DOI":"10.1016\/j.jpeds.2017.06.042","article-title":"Presymptomatic Diagnosis of Spinal Muscular Atrophy Through Newborn Screening","volume":"190","author":"Chien","year":"2017","journal-title":"J. Pediatr."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"343","DOI":"10.1016\/j.nmd.2019.02.003","article-title":"Newborn screening for SMA in Southern Belgium","volume":"29","author":"Boemer","year":"2019","journal-title":"Neuromuscul. Disord."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1255","DOI":"10.3389\/fgene.2019.01255","article-title":"Newborn Screening for Spinal Muscular Atrophy in China Using DNA Mass Spectrometry","volume":"10","author":"Lin","year":"2019","journal-title":"Front. Genet."},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Shinohara, M., Niba, E.T.E., Wijaya, Y.O.S., Takayama, I., Mitsuishi, C., Kumasaka, S., Kondo, Y., Takatera, A., Hokuto, I., and Morioka, I. (2019). A novel system for spinal muscular atrophy screening in newborns: Japanese pilot study. Int. J. Neonatal Screen., 5.","DOI":"10.3390\/ijns5040041"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"503","DOI":"10.3233\/JND-190428","article-title":"One year of newborn screening for SMA\u2013Results of a German pilot project","volume":"6","author":"Vill","year":"2019","journal-title":"J. Neuromuscul. Dis."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"23","DOI":"10.1038\/s41431-019-0476-4","article-title":"High-throughput genetic newborn screening for spinal muscular atrophy by rapid nucleic acid extraction from dried blood spots and 384-well qPCR","volume":"28","author":"Czibere","year":"2020","journal-title":"Eur. J. Hum. Genet."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1296","DOI":"10.1038\/s41436-020-0824-3","article-title":"Implementation of population-based newborn screening reveals low incidence of spinal muscular atrophy","volume":"22","author":"Kay","year":"2020","journal-title":"Genet. Med."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"111","DOI":"10.4236\/ojgen.2013.32014","article-title":"1000 sample comparison of MLPA and RT-PCR for carrier detection and diagnostic testing for Spinal Muscular Atrophy Type 1","volume":"3","author":"Strom","year":"2013","journal-title":"Open J. Genet."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"101","DOI":"10.3343\/alm.2021.41.1.101","article-title":"Detection of Spinal Muscular Atrophy Using a Duplexed Real-Time PCR Approach With Locked Nucleic Acid-Modified Primers","volume":"41","author":"Pan","year":"2021","journal-title":"Ann. Lab. Med."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"428","DOI":"10.1097\/01.gim.0000227970.60450.b2","article-title":"A feasibility study for the newborn screening of spinal muscular atrophy","volume":"8","author":"Pyatt","year":"2006","journal-title":"Genet. Med."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"74","DOI":"10.1002\/humu.10221","article-title":"Determination of SMN1 and SMN2 copy number using TaqManTM technology","volume":"22","author":"Anhuf","year":"2003","journal-title":"Hum. Mutat."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"37","DOI":"10.1089\/gtmb.2008.0084","article-title":"A Simple Multiplex Real-Time PCR Methodology for the SMN1 Gene Copy Number Quantification","volume":"13","author":"Passon","year":"2009","journal-title":"Genet. Test. Mol. Biomark."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"504","DOI":"10.1017\/cjn.2020.229","article-title":"Newborn Screening for Spinal Muscular Atrophy: Ontario Testing and Follow-up Recommendations","volume":"48","author":"McMillan","year":"2020","journal-title":"Can. J. Neurol. Sci. J. Can. Sci. Neurol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1340","DOI":"10.1086\/302369","article-title":"Quantitative analysis of survival motor neuron copies: Identification of subtle SMN1 mutations in patients with spinal muscular atrophy, genotype- phenotype correlation, and implications for genetic counseling","volume":"64","author":"Wirth","year":"1999","journal-title":"Am. J. Hum. Genet."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1608","DOI":"10.1002\/ajmg.a.33474","article-title":"Newborn and carrier screening for spinal muscular atrophy","volume":"152","author":"Prior","year":"2010","journal-title":"Am. J. Med. Genet. Part A"},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Strunk, A., Abbes, A., Stuitje, A.R., Hettinga, C., Sepers, E.M., Snetselaar, R., Schouten, J., Asselman, F.L., Cuppen, I., and Lemmink, H. (2019). Validation of a fast, robust, inexpensive, two-tiered neonatal screening test algorithm on dried blood spots for spinal muscular atrophy. Int. J. Neonatal Screen., 5.","DOI":"10.3390\/ijns5020021"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"608","DOI":"10.1038\/gim.2017.152","article-title":"Pilot study of population-based newborn screening for spinal muscular atrophy in New York state","volume":"20","author":"Kraszewski","year":"2018","journal-title":"Genet. Med."},{"key":"ref_34","unstructured":"RCoreTeam (2021, July 03). R: A Language and Environment for Statistical Computing; R Foundation for Statistical Computing: Vienna, Austria. Available online: https:\/\/www.R-project.org\/."},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Gutierrez-Mateo, C., Timonen, A., Vaahtera, K., Jaakkola, M., Hougaard, D.M., Bybjerg-Grauholm, J., Baekvad-Hansen, M., Adamsen, D., Filippov, G., and Dallaire, S. (2019). Development of a multiplex real-time PCR assay for the newborn screening of SCID, SMA, and XLA. Int. J. Neonatal Screen., 5.","DOI":"10.3390\/ijns5040039"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"569","DOI":"10.1089\/gtmb.2020.0085","article-title":"Intelligent Ratio: A New Method for Carrier and Newborn Screening in Spinal Muscular Atrophy","volume":"24","author":"Cavdarli","year":"2020","journal-title":"Genet. Test. Mol. Biomark."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"217","DOI":"10.1053\/j.semperi.2015.03.006","article-title":"Newborn screening for spinal muscular atrophy: Anticipating an imminent need","volume":"39","author":"Phan","year":"2015","journal-title":"Semin. Perinatol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1753","DOI":"10.1373\/clinchem.2018.293712","article-title":"Multiplex droplet digital PCR method applicable to newborn screening, carrier status, and assessment of spinal muscular atrophy","volume":"64","author":"Gavrilov","year":"2018","journal-title":"Clin. Chem."},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Kellar-Guenther, Y., McKasson, S., Hale, K., Singh, S., Sontag, M.K., and Ojodu, J. (2020). Implementing statewide newborn screening for new disorders: U.S. Program experiences. Int. J. Neonatal Screen., 6.","DOI":"10.3390\/ijns6020035"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"97","DOI":"10.3233\/JND-190468","article-title":"Revised Recommendations for the Treatment of Infants Diagnosed with Spinal Muscular Atrophy Via Newborn Screening Who Have 4 Copies of SMN2","volume":"7","author":"Glascock","year":"2020","journal-title":"J. Neuromuscul. Dis."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"109","DOI":"10.3233\/JND-200475","article-title":"Infants Diagnosed with Spinal Muscular Atrophy and 4 SMN2 Copies through Newborn Screening-Opportunity or Burden?","volume":"7","author":"Vill","year":"2020","journal-title":"J. Neuromuscul. Dis."},{"key":"ref_42","unstructured":"Kemper, A.R., Lam, K., Comeau, A.M., Kwon, J., Green, N.S., Ojodu, J., Grosse, S., Prosser, L.A., Jones, E., and Tanksley, S. (2018). Evidence-Based Review of Newborn Screening for Spinal Muscular Atrophy (SMA): Final Report (v5.2), United States Secretary of Health and Human Services Advisory Committee on Heritable Disorders in Newborns and Children, Maternal Child Health Bureau."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"1153","DOI":"10.2147\/TCRM.S172291","article-title":"Clinical evidence supporting early treatment of patients with spinal muscular atrophy: Current perspectives","volume":"15","author":"Dangouloff","year":"2019","journal-title":"Ther. Clin. Risk Manag."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"157","DOI":"10.1002\/mus.24497","article-title":"Spinal muscular atrophy: Diagnosis and management in a new therapeutic era","volume":"51","author":"Arnold","year":"2015","journal-title":"Muscle Nerve"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"11","DOI":"10.2147\/TACG.S239603","article-title":"Spinal muscular atrophy: Mutations, testing, and clinical relevance","volume":"14","author":"Keinath","year":"2021","journal-title":"Appl. Clin. Genet."}],"container-title":["International Journal of Neonatal Screening"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2409-515X\/7\/3\/53\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T06:43:52Z","timestamp":1760165032000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2409-515X\/7\/3\/53"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,8,11]]},"references-count":45,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2021,9]]}},"alternative-id":["ijns7030053"],"URL":"https:\/\/doi.org\/10.3390\/ijns7030053","relation":{},"ISSN":["2409-515X"],"issn-type":[{"value":"2409-515X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,8,11]]}}}