{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,31]],"date-time":"2026-07-31T02:54:33Z","timestamp":1785466473957,"version":"3.56.0"},"reference-count":31,"publisher":"Oxford University Press (OUP)","issue":"7","license":[{"start":{"date-parts":[[2021,4,6]],"date-time":"2021-04-06T00:00:00Z","timestamp":1617667200000},"content-version":"vor","delay-in-days":1,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"name":"Special Research Funding"},{"DOI":"10.13039\/501100004385","name":"Ghent University","doi-asserted-by":"publisher","award":["BOF18\/DOC\/200"],"award-info":[{"award-number":["BOF18\/DOC\/200"]}],"id":[{"id":"10.13039\/501100004385","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Flemish Fund for Scientific Research","award":["G051017N"],"award-info":[{"award-number":["G051017N"]}]},{"name":"Flemish Fund for Scientific Research","award":["G051516N"],"award-info":[{"award-number":["G051516N"]}]},{"name":"Flemish Fund for Scientific Research","award":["G1507816N"],"award-info":[{"award-number":["G1507816N"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2021,7,6]]},"abstract":"<jats:title>Abstract<\/jats:title>\n               <jats:sec>\n                  <jats:title>Background<\/jats:title>\n                  <jats:p>The quantification of mitochondrial DNA heteroplasmy for the diagnosis of mitochondrial disease or after mitochondrial donation, is performed mainly using next-generation sequencing strategies (NGS). Digital PCR (dPCR) has the potential to offer an accurate alternative for mutation load quantification.<\/jats:p>\n               <\/jats:sec>\n               <jats:sec>\n                  <jats:title>Methods<\/jats:title>\n                  <jats:p>We assessed the mutation load of 23 low-input human samples at the m.11778 locus, which is associated with Leber\u2019s hereditary optic neuropathy (LHON) using 2 droplet digital PCR platforms (Stilla Naica and Bio-Rad QX200) and the standard NGS strategy. Assay validation was performed by analyzing a titration series with mutation loads ranging from 50% to 0.01%.<\/jats:p>\n               <\/jats:sec>\n               <jats:sec>\n                  <jats:title>Results<\/jats:title>\n                  <jats:p>A good concordance in mutation rates was observed between both dPCR techniques and NGS. dPCR established a distinctly lower level of background noise compared to NGS. Minor alleles with mutation loads lower than 1% could still be detected, with standard deviations of the technical replicates varying between 0.07% and 0.44% mutation load. Although no significant systematic bias was observed when comparing dPCR and NGS, a minor proportional bias was detected. A slight overestimation of the minor allele was observed for the NGS data, most probably due to amplification and sequencing errors in the NGS workflow.<\/jats:p>\n               <\/jats:sec>\n               <jats:sec>\n                  <jats:title>Conclusion<\/jats:title>\n                  <jats:p>dPCR has proven to be an accurate tool for the quantification of mitochondrial heteroplasmy, even for samples harboring a low mutation load (&amp;lt;1%). In addition, this alternative technique holds multiple benefits compared to NGS (e.g., less hands-on time, more straightforward data-analysis, and a lower up-front capital investment).<\/jats:p>\n               <\/jats:sec>","DOI":"10.1093\/clinchem\/hvab021","type":"journal-article","created":{"date-parts":[[2021,2,3]],"date-time":"2021-02-03T06:09:23Z","timestamp":1612332563000},"page":"968-976","source":"Crossref","is-referenced-by-count":8,"title":["Digital Polymerase Chain Reaction for Assessment of Mutant Mitochondrial Carry-over after Nuclear Transfer for In Vitro Fertilization"],"prefix":"10.1093","volume":"67","author":[{"given":"Olivier","family":"Tytgat","sequence":"first","affiliation":[{"name":"Laboratory of Pharmaceutical Biotechnology, Ghent University, Ghent, Belgium"},{"name":"Department of Life Science Technologies, Imec, Leuven, 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