{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,29]],"date-time":"2026-09-29T18:26:15Z","timestamp":1790706375724,"version":"4.1.0"},"reference-count":20,"publisher":"Oxford University Press (OUP)","issue":"8","license":[{"start":{"date-parts":[[2020,7,24]],"date-time":"2020-07-24T00:00:00Z","timestamp":1595548800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100000024","name":"Canadian Institute for Health Research","doi-asserted-by":"crossref","id":[{"id":"10.13039\/501100000024","id-type":"DOI","asserted-by":"crossref"}]},{"name":"Canadian Foundation for Innovation, Genome Canada and British Columbia"},{"name":"Mathematics of Information Technology and Complex Systems"},{"name":"PROOF Centre of Excellence"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2020,8,1]]},"abstract":"<jats:title>Abstract<\/jats:title>\n               <jats:sec>\n                  <jats:title>Background<\/jats:title>\n                  <jats:p>HEARTBiT is a whole blood-based gene profiling assay using the nucleic acid counting NanoString technology for the exclusionary diagnosis of acute cellular rejection in heart transplant patients. The HEARTBiT score measures the risk of acute cellular rejection in the first year following heart transplant, distinguishing patients with stable grafts from those at risk for acute cellular rejection. Here, we provide the analytical performance characteristics of the HEARTBiT assay and the results on pilot clinical validation.<\/jats:p>\n               <\/jats:sec>\n               <jats:sec>\n                  <jats:title>Methods<\/jats:title>\n                  <jats:p>We used purified RNA collected from PAXgene blood samples to evaluate the characteristics of a 12-gene panel HEARTBiT assay, for its linearity range, quantitative bias, precision, and reproducibility. These parameters were estimated either from serial dilutions of individual samples or from repeated runs on pooled samples.<\/jats:p>\n               <\/jats:sec>\n               <jats:sec>\n                  <jats:title>Results<\/jats:title>\n                  <jats:p>We found that all 12 genes showed linear behavior within the recommended assay input range of 125\u2009ng to 500\u2009ng of purified RNA, with most genes showing 3% or lower quantitative bias and around 5% coefficient of variation. Total variation resulting from unique operators, reagent lots, and runs was less than 0.02 units standard deviation (SD). The performance of the analytically validated assay (AUC\u2009=\u20090.75) was equivalent to what we observed in the signature development dataset.<\/jats:p>\n               <\/jats:sec>\n               <jats:sec>\n                  <jats:title>Conclusion<\/jats:title>\n                  <jats:p>The analytical performance of the assay within the specification input range demonstrated reliable quantification of the HEARTBiT score within 0.02 SD units, measured on a 0 to 1 unit scale. This assay may therefore be of high utility in clinical validation of HEARTBiT in future biomarker observational trials.<\/jats:p>\n               <\/jats:sec>","DOI":"10.1093\/clinchem\/hvaa123","type":"journal-article","created":{"date-parts":[[2020,6,6]],"date-time":"2020-06-06T11:10:21Z","timestamp":1591441821000},"page":"1063-1071","source":"Crossref","is-referenced-by-count":8,"title":["Analytical Validation of HEARTBiT: A Blood-Based Multiplex Gene Expression Profiling Assay for Exclusionary Diagnosis of Acute Cellular Rejection in Heart Transplant Patients"],"prefix":"10.1093","volume":"66","author":[{"given":"Ji-Young V","family":"Kim","sequence":"first","affiliation":[{"name":"Prevention of Organ Failure (PROOF) Centre of Excellence, Vancouver, BC, Canada"},{"name":"Centre for Heart Lung Innovation, St Paul\u2019s Hospital, University of British Columbia, Vancouver, BC, Canada"},{"name":"Division of Respiratory Medicine, Department of Medicine, University of British Columbia, Vancouver, BC, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Brandon","family":"Lee","sequence":"additional","affiliation":[{"name":"Prevention of Organ Failure (PROOF) Centre of Excellence, Vancouver, BC, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pavlos","family":"Koitsopoulos","sequence":"additional","affiliation":[{"name":"Department of Medicine, University of British Columbia, Vancouver, BC, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Casey P","family":"Shannon","sequence":"additional","affiliation":[{"name":"Prevention of Organ Failure (PROOF) Centre of Excellence, Vancouver, BC, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Virginia","family":"Chen","sequence":"additional","affiliation":[{"name":"Prevention of Organ Failure (PROOF) Centre of Excellence, Vancouver, BC, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zsuzsanna","family":"Hollander","sequence":"additional","affiliation":[{"name":"Prevention of Organ Failure (PROOF) Centre of Excellence, Vancouver, BC, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sara","family":"Assadian","sequence":"additional","affiliation":[{"name":"Prevention of Organ Failure (PROOF) Centre of Excellence, Vancouver, BC, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Karen","family":"Lam","sequence":"additional","affiliation":[{"name":"Prevention of Organ Failure (PROOF) Centre of Excellence, Vancouver, BC, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gordon","family":"Ritchie","sequence":"additional","affiliation":[{"name":"Department of Pathology & Laboratory Medicine, University of British Columbia, Vancouver, BC, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Janet","family":"McManus","sequence":"additional","affiliation":[{"name":"netCAD Canadian Blood Services, Vancouver, BC, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"W Robert","family":"McMaster","sequence":"additional","affiliation":[{"name":"Department of Medical Genetics, University of British Columbia, Vancouver, BC, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Raymond T","family":"Ng","sequence":"additional","affiliation":[{"name":"Department of Computer Science, University of British Columbia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bruce M","family":"McManus","sequence":"additional","affiliation":[{"name":"Prevention of Organ Failure (PROOF) Centre of Excellence, Vancouver, BC, Canada"},{"name":"Department of Pathology & Laboratory Medicine, University of British Columbia, Vancouver, BC, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Scott J","family":"Tebbutt","sequence":"additional","affiliation":[{"name":"Prevention of Organ Failure (PROOF) Centre of Excellence, Vancouver, BC, Canada"},{"name":"Centre for Heart Lung Innovation, St Paul\u2019s Hospital, University of British Columbia, Vancouver, BC, Canada"},{"name":"Division of Respiratory Medicine, Department of Medicine, University of British Columbia, Vancouver, BC, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2020,7,24]]},"reference":[{"key":"2020080407304400800_hvaa123-B1","doi-asserted-by":"crossref","first-page":"1089","DOI":"10.1016\/j.healun.2010.08.007","article-title":"The Registry of the International Society for Heart and Lung Transplantation: twenty-seventh official adult heart transplant report\u20132010","volume":"29","author":"Stehlik","year":"2010","journal-title":"J Heart Lung Transplant"},{"key":"2020080407304400800_hvaa123-B2","doi-asserted-by":"crossref","first-page":"584","DOI":"10.1016\/j.cgh.2016.07.035","article-title":"Acute rejection increases risk of graft failure and death in recent liver transplant recipients","volume":"15","author":"Levitsky","year":"2017","journal-title":"Clin Gastroenterol Hepatol"},{"key":"2020080407304400800_hvaa123-B3","doi-asserted-by":"crossref","first-page":"914","DOI":"10.1016\/j.healun.2010.05.034","article-title":"The International Society of Heart and Lung Transplantation Guidelines for the care of heart transplant recipients","volume":"29","author":"Costanzo","year":"2010","journal-title":"J Heart Lung Transplant"},{"key":"2020080407304400800_hvaa123-B4","first-page":"239","article-title":"Reproducibility of the acute rejection diagnosis in human cardiac allografts. The Stanford Classification and the International Grading System","volume":"12","author":"Nielsen","year":"1993","journal-title":"J Heart Lung Transplant"},{"key":"2020080407304400800_hvaa123-B5","doi-asserted-by":"crossref","first-page":"1172","DOI":"10.1097\/TP.0b013e31826e19e2","article-title":"Concordance among pathologists in the second cardiac allograft rejection gene expression observational study (CARGO II)","volume":"94","author":"Crespo-Leiro","year":"2012","journal-title":"Transplantation"},{"key":"2020080407304400800_hvaa123-B6","first-page":"55","article-title":"Endomyocardial biopsy for monitoring heart transplant patients: 11-years-experience at a German heart center","volume":"6","author":"Strecker","year":"2013","journal-title":"Int J Clin Exp Pathol"},{"key":"2020080407304400800_hvaa123-B7","doi-asserted-by":"crossref","first-page":"83784","DOI":"10.18632\/oncotarget.11965","article-title":"Nationwide population-based study reveals increased malignancy risk in Taiwanese liver transplant recipients","volume":"7","author":"Tsai","year":"2016","journal-title":"Oncotarget"},{"key":"2020080407304400800_hvaa123-B8","doi-asserted-by":"crossref","first-page":"76","DOI":"10.3389\/fmolb.2018.00076","article-title":"Molecular diagnostics in clinical oncology","volume":"5","author":"Sokolenko","year":"2018","journal-title":"Front Mol Biosci"},{"key":"2020080407304400800_hvaa123-B9","article-title":"HEARTBiT: a transcriptomic signature for excluding acute cellular rejection in adult heart allograft patients","author":"Shannon","journal-title":"Can J Cardiol"},{"key":"2020080407304400800_hvaa123-B10","doi-asserted-by":"crossref","first-page":"317","DOI":"10.1038\/nbt1385","article-title":"Direct multiplexed measurement of gene expression with color-coded probe pairs","volume":"26","author":"Geiss","year":"2008","journal-title":"Nat Biotechnol"},{"key":"2020080407304400800_hvaa123-B11","first-page":"11","article-title":"Evaluation of the linearity of quantitative measurement procedures: a statistical approach; approved guideline","volume":"23","year":"2003","journal-title":"CLSI\/NCCLS Document EP6-A"},{"key":"2020080407304400800_hvaa123-B12","year":"2019"},{"key":"2020080407304400800_hvaa123-B13","doi-asserted-by":"crossref","first-page":"1084","DOI":"10.1373\/clinchem.2006.076497","article-title":"Analytical validation of the Oncotype DX genomic diagnostic test for recurrence prognosis and therapeutic response prediction in node-negative, estrogen receptor-positive breast cancer","volume":"53","author":"Cronin","year":"2007","journal-title":"Clin Chem"},{"key":"2020080407304400800_hvaa123-B14","doi-asserted-by":"crossref","first-page":"177","DOI":"10.1186\/1471-2407-14-177","article-title":"Analytical validation of the PAM50-based Prosigna Breast Cancer Prognostic Gene Signature Assay and nCounter Analysis System using formalin-fixed paraffin-embedded breast tumor specimens","volume":"14","author":"Nielsen","year":"2014","journal-title":"BMC Cancer"},{"key":"2020080407304400800_hvaa123-B15","doi-asserted-by":"crossref","first-page":"355","DOI":"10.1177\/1948550617697177","article-title":"Equivalence tests: a practical primer for t tests, correlations, and meta-analyses","volume":"8","author":"Lakens","year":"2017","journal-title":"Soc Psychol Personal Sci"},{"key":"2020080407304400800_hvaa123-B16","doi-asserted-by":"crossref","first-page":"1","DOI":"10.18637\/jss.v067.i01","article-title":"Fitting linear mixed-effects models using lme4","volume":"67","author":"Bates","year":"2015","journal-title":"J Stat Softw"},{"key":"2020080407304400800_hvaa123-B17","doi-asserted-by":"crossref","first-page":"1857","DOI":"10.1016\/S0041-1345(02)03072-5","article-title":"Is there a need for surveillance endomyocardial biopsies in heart transplantation?","volume":"34","author":"Kirklin","year":"2002","journal-title":"Transplant"},{"key":"2020080407304400800_hvaa123-B18","doi-asserted-by":"crossref","first-page":"775","DOI":"10.1016\/j.healun.2014.04.019","article-title":"A contemporary review of adult heart transplantation: 2012 to 2013","volume":"33","author":"DePasquale","year":"2014","journal-title":"J Heart Lung Transplant"},{"key":"2020080407304400800_hvaa123-B19","doi-asserted-by":"crossref","first-page":"1890","DOI":"10.1056\/NEJMoa0912965","article-title":"Gene-expression profiling for rejection surveillance after cardiac transplantation","volume":"362","author":"Pham","year":"2010","journal-title":"N Engl J Med"},{"key":"2020080407304400800_hvaa123-B20","year":"2019"}],"container-title":["Clinical Chemistry"],"original-title":[],"language":"en","link":[{"URL":"http:\/\/academic.oup.com\/clinchem\/advance-article-pdf\/doi\/10.1093\/clinchem\/hvaa123\/33527484\/hvaa123.pdf","content-type":"application\/pdf","content-version":"am","intended-application":"syndication"},{"URL":"http:\/\/academic.oup.com\/clinchem\/article-pdf\/66\/8\/1063\/33563166\/hvaa123.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"http:\/\/academic.oup.com\/clinchem\/article-pdf\/66\/8\/1063\/33563166\/hvaa123.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2020,8,4]],"date-time":"2020-08-04T11:32:40Z","timestamp":1596540760000},"score":1,"resource":{"primary":{"URL":"https:\/\/academic.oup.com\/clinchem\/article\/66\/8\/1063\/5875790"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,7,24]]},"references-count":20,"journal-issue":{"issue":"8","published-online":{"date-parts":[[2020,7,24]]},"published-print":{"date-parts":[[2020,8,1]]}},"URL":"https:\/\/doi.org\/10.1093\/clinchem\/hvaa123","relation":{},"ISSN":["0009-9147","1530-8561"],"issn-type":[{"value":"0009-9147","type":"print"},{"value":"1530-8561","type":"electronic"}],"subject":[],"published-other":{"date-parts":[[2020,8]]},"published":{"date-parts":[[2020,7,24]]}}}