{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,23]],"date-time":"2026-10-23T19:47:50Z","timestamp":1792784870203,"version":"4.5.0"},"reference-count":34,"publisher":"Springer Science and Business Media LLC","license":[{"start":{"date-parts":[[2026,9,22]],"date-time":"2026-09-22T00:00:00Z","timestamp":1790035200000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2026,9,22]],"date-time":"2026-09-22T00:00:00Z","timestamp":1790035200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"name":"MRC Holland b.v.","id":[{"id":"https:\/\/ror.org\/02wcat891","id-type":"ROR","asserted-by":"crossref"}]}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Eur J Hum Genet"],"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    Decreasing DNA sequencing costs has paved the way to the use of next generation sequencing (NGS) to expand the number of conditions tested in routine newborn screening (NBS) programs. Expanding NBS programs by NGS, however, raises technical, ethical and economic concerns that are important for low-middle income countries or countries that are hesitant to use sequencing in a public health setting. qPCR-based DNA analyses have already been implemented in routine NBS programs; however, qPCR cannot be highly multiplexed. digitalMLPA (dMLPA) is a highly multiplexed probe-based DNA technique. Here, we describe the new dMLPA EZtec-MS technique that was specifically developed for use on dried blot spots (DBS) and can target up to 1200 DNA sequences representing different variant types, allowing high-throughput testing of crude DNA extracts prepared from a single DBS punch. The assay was validated using positive DNA samples and crude DNA extracts from DBS of 2069 negatively screened Philippine newborns. The EZtec-MS assay used in this study successfully detected copy number variants, (including complex genomic regions (\n                    <jats:italic>SMN1\/2<\/jats:italic>\n                    ,\n                    <jats:italic>HBA1\/2<\/jats:italic>\n                    )), inversions (\n                    <jats:italic>F8<\/jats:italic>\n                    ), methylation (imprinting diseases, Fragile X), single nucleotide variants (including\n                    <jats:italic>ATP7B, CTNS<\/jats:italic>\n                    ,\n                    <jats:italic>MT-RNR1)<\/jats:italic>\n                    , and low copy number sequences (TRECs for SCID, cytomegalovirus) in a single reaction in both positive DNA and crude DNA extracts from DBS. The results also demonstrated that dMLPA EZtec-MS is a high-throughput method, suitable for DBS, and has a relatively short turn-around time of 36\u201340\u2009h. Our findings indicate that the dMLPA EZtec-MS technique is a promising versatile tool for DNA-based NBS programs.\n                  <\/jats:p>","DOI":"10.1038\/s41431-026-02235-2","type":"journal-article","created":{"date-parts":[[2026,9,22]],"date-time":"2026-09-22T07:08:17Z","timestamp":1790060897000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["digitalMLPA EZtec-MS: A new highly multiplexed DNA probe-based technique for neonatal screening"],"prefix":"10.1038","author":[{"ORCID":"https:\/\/orcid.org\/0009-0002-3244-301X","authenticated-orcid":false,"given":"Terence Diane","family":"Fabella","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6614-876X","authenticated-orcid":false,"given":"Joery","family":"den Hoed","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eva Maria","family":"Cutiongco-de la Paz","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Carmencita","family":"Padilla","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chris","family":"Hettinga","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4554-9796","authenticated-orcid":false,"given":"Karel","family":"de Groot","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Martin","family":"Lod\u00e9n-van Straaten","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0000-5188-1168","authenticated-orcid":false,"given":"Tatiana C.","family":"Santos","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Richiel","family":"van den Berg","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7187-4563","authenticated-orcid":false,"given":"Erik A.","family":"Sistermans","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0006-8747-5085","authenticated-orcid":false,"given":"Jan","family":"Schouten","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2026,9,22]]},"reference":[{"key":"2235_CR1","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1001\/jamanetworkopen.2025.38198","volume":"8","author":"D Carli","year":"2025","unstructured":"Carli D, Quarello P, Porta F, Cagnazzo C, Zucchetti G, Proto CF, et al. A Genomic Sequencing Approach to Newborn Mass Screening and Its Opportunities. JAMA Netw Open. 2025;8:1\u201312.","journal-title":"JAMA Netw Open"},{"key":"2235_CR2","doi-asserted-by":"publisher","first-page":"21","DOI":"10.3390\/ijns5020021","volume":"5","author":"A Strunk","year":"2019","unstructured":"Strunk A, Abbes A, Stuitje AR, Hettinga C, Sepers EM, Snetselaar R, et al. Validation of a Fast, Robust, Inexpensive, Two-Tiered Neonatal Screening Test algorithm on Dried Blood Spots for Spinal Muscular Atrophy. Int J Neonatal Screen. 2019;5:21.","journal-title":"Int J Neonatal Screen"},{"key":"2235_CR3","doi-asserted-by":"crossref","unstructured":"Brku\u0161anin M, Garai N, Karanovi\u0107 J, \u0160ljivan\u010danin Jakovljevi\u0107 T, Dimitrijevi\u0107 A, Jovanovi\u0107 K et al. Our Journey from Individual Efforts to Nationwide Support: Implementing Newborn Screening for Spinal Muscular Atrophy in Serbia. 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J.S. is the owner and CEO, and J.d.H., C.H., K.D.G., M.L.S., R.v.d.B., and T.S. are employees of MRC Holland. MRC Holland is a company that developed and manufactures the molecular diagnostic test MLPA. Other authors declare no conflicts of interest.","order":1,"name":"Ethics","label":"Competing interests","group":{"name":"EthicsHeading","label":"Ethics"}},{"value":"The project was approved by the University of the Philippines Manila, Research Ethics Board (UPMREB 2023-0224-EX). The Philippine Republic Act (RA) 10173 or Data Privacy Act of 2012 establishes the rule that anonymized or de-identified samples are not personal information. Informed consent was therefore not required for DBS use.","order":2,"name":"Ethics","label":"Ethical approval","group":{"name":"EthicsHeading","label":"Ethics"}}]}}