{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,7]],"date-time":"2026-07-07T21:46:05Z","timestamp":1783460765285,"version":"3.55.0"},"reference-count":76,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2026,3,26]],"date-time":"2026-03-26T00:00:00Z","timestamp":1774483200000},"content-version":"vor","delay-in-days":84,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,1,1]],"date-time":"2026-01-01T00:00:00Z","timestamp":1767225600000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Journal of Diabetes Research"],"published-print":{"date-parts":[[2026,1]]},"abstract":"<jats:sec>\n                    <jats:title>Background<\/jats:title>\n                    <jats:p>Mitochondrial DNA m.3243A&gt;G variant causes maternally inherited diabetes with deafness. We developed a droplet digital PCR (dPCR) assay to quantify m.3243A&gt;G heteroplasmy and investigated its prevalence in Chinese Han early\u2010onset diabetes patients.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      The dPCR method was validated using plasmid mixtures (10\n                      <jats:sup>4<\/jats:sup>\n                      copies) for sensitivity, specificity, dynamic range (10\u201310\n                      <jats:sup>5<\/jats:sup>\n                      copies\/\u03bcL), accuracy, and precision. We screened 1506 unselected early\u2010onset diabetes patients, analyzing blood and urinary sediment DNA. This population comprised 1474 diagnosed type 2 diabetes patients (97.88%), 31 diagnosed type 1 diabetes patients (2.06%), and one patient with mitochondrial diabetes (0.07%). The dPCR results were compared with Sanger sequencing and next generation sequencing (NGS) to evaluate the accuracy of heteroplasmy rate quantification.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      The dynamic range spans from 10 to 10\n                      <jats:sup>5<\/jats:sup>\n                      copies\/\u03bcL. The assay detected heteroplasmy as low as 0.05% with 100% specificity. The method can reliably quantify the heteroplasmy rate at 0% to 100% for an input copy number of 10\n                      <jats:sup>4<\/jats:sup>\n                      . Using the dPCR method, we identified 0.80% (\n                      <jats:italic>n<\/jats:italic>\n                      \u2009=\u200912) had &gt;5% m.3243A&gt;G heteroplasmy in peripheral blood and\/or urinary sediment, and 0.40% (\n                      <jats:italic>n<\/jats:italic>\n                      \u2009=\u20096) had 1%\u20135% among the early\u2010onset diabetes patients. The dPCR approach was more sensitive than Sanger sequencing. When compared with NGS results, the dPCR method accurately genotyped m.3243A&gt;G and quantified its heteroplasmy rate in clinical samples. Furthermore, it demonstrated that urine is a reliable noninvasive sample source for determining the m.3243A&gt;G heteroplasmy rate.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>The developed method can provide an accurate, reliable, rapid, simple, and low\u2010cost evaluation of the heteroplasmy rate of m.3243A&gt;G for both peripheral blood and urinary sediment. Thus, it is highly promising as a genetic screening method for early\u2010onset diabetes related to m.3243A&gt;G.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1155\/jdr\/9298888","type":"journal-article","created":{"date-parts":[[2026,3,26]],"date-time":"2026-03-26T23:24:58Z","timestamp":1774567498000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Clinical Validation of Digital PCR for Precise m.3243A&gt;G Heteroplasmy Quantification in Early\u2010Onset 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