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However, a higher-order multiplexing dPCR assay for measuring\n                    <jats:italic>SMN1<\/jats:italic>\n                    and\n                    <jats:italic>SMN2<\/jats:italic>\n                    copy numbers in spinal muscular atrophy (SMA) samples has not been reported. Described here is a rapid multiplex SMA dPCR genotyping assay run on a fully integrated dPCR instrument with five optical channels. The hydrolysis probe-based multiplex dPCR assay quantifies\n                    <jats:italic>SMN1<\/jats:italic>\n                    ,\n                    <jats:italic>SMN2,<\/jats:italic>\n                    and the total\n                    <jats:italic>SMN<\/jats:italic>\n                    (\n                    <jats:italic>SMN1<\/jats:italic>\n                    \u2009+\u2009\n                    <jats:italic>SMN2<\/jats:italic>\n                    ) while using\n                    <jats:italic>RPPH1<\/jats:italic>\n                    gene as an internal reference control. The quadruplex assay was evaluated with characterized control DNA samples and validated with 15 blinded clinical samples from a previously published study.\n                    <jats:italic>SMN1<\/jats:italic>\n                    and\n                    <jats:italic>SMN2<\/jats:italic>\n                    copy numbers were completely concordant with previous results for both the control and blinded samples. The dPCR-based SMA copy number determination was accomplished in 90\u00a0min with a walk-away workflow identical to real-time quantitative PCR (qPCR). In summary, presented here is a simple higher-order multiplexing solution on a novel digital PCR platform to meet the growing demand for SMA genotyping and prognostics.\n                  <\/jats:p>","DOI":"10.1038\/s41598-020-76893-7","type":"journal-article","created":{"date-parts":[[2020,11,16]],"date-time":"2020-11-16T06:04:49Z","timestamp":1605506689000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":25,"title":["Development and validation of a 4-color multiplexing spinal muscular atrophy (SMA) genotyping assay on a novel integrated digital PCR instrument"],"prefix":"10.1038","volume":"10","author":[{"given":"Lingxia","family":"Jiang","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Robert","family":"Lin","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Steve","family":"Gallagher","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andrew","family":"Zayac","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Matthew E. 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