{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,15]],"date-time":"2026-04-15T13:54:51Z","timestamp":1776261291155,"version":"3.50.1"},"reference-count":52,"publisher":"MDPI AG","issue":"1","license":[{"start":{"date-parts":[[2022,1,5]],"date-time":"2022-01-05T00:00:00Z","timestamp":1641340800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Diagnostics"],"abstract":"<jats:p>The use of droplet digital PCR (ddPCR) to identify and quantify low-abundance targets is a significant advantage for accurately detecting potentially oncogenic bacteria. Fusobacterium nucleatum (Fn) is implicated in colorectal cancer (CRC) tumorigenesis and is becoming an important prognostic biomarker. We evaluated the detection accuracy and clinical relevance of Fn DNA by ddPCR in a molecularly characterized, formalin-fixed, paraffin-embedded (FFPE) CRC cohort previously analyzed by qPCR for Fn levels. Following a ddPCR assay optimization and an analytical evaluation, Fn DNA were measured in 139 CRC FFPE cases. The measures of accuracy for Fn status compared to the prior results generated by qPCR and the association with clinicopathological and molecular patients\u2019 features were also evaluated. The ddPCR-based Fn assay was sensitive and specific to positive controls. Fn DNA were detected in 20.1% of cases and further classified as Fn-high and Fn-low\/negative, according to the median amount of Fn DNA that were detected in all cases and associated with the patient\u2019s worst prognosis. There was a low agreement between the Fn status determined by ddPCR and qPCR (Cohen\u2019s Kappa = 0.210). Our findings show that ddPCR can detect and quantify Fn in FFPE tumor tissues and highlights its clinical relevance in Fn detection in a routine CRC setting.<\/jats:p>","DOI":"10.3390\/diagnostics12010114","type":"journal-article","created":{"date-parts":[[2022,1,5]],"date-time":"2022-01-05T20:42:16Z","timestamp":1641415336000},"page":"114","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":13,"title":["Accuracy and Clinical Relevance of Intra-Tumoral Fusobacterium nucleatum Detection in Formalin-Fixed Paraffin-Embedded (FFPE) Tissue by Droplet Digital PCR (ddPCR) in Colorectal Cancer"],"prefix":"10.3390","volume":"12","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-9682-7517","authenticated-orcid":false,"given":"Jos\u00e9 Guilherme","family":"Datorre","sequence":"first","affiliation":[{"name":"Molecular Oncology Research Center, Barretos Cancer Hospital, Barretos 14784400, Brazil"}]},{"given":"Ana Carolina","family":"de Carvalho","sequence":"additional","affiliation":[{"name":"Molecular Oncology Research Center, Barretos Cancer Hospital, Barretos 14784400, Brazil"}]},{"given":"Mariana Bisarro","family":"dos Reis","sequence":"additional","affiliation":[{"name":"Molecular Oncology Research Center, Barretos Cancer Hospital, Barretos 14784400, Brazil"}]},{"given":"Monise","family":"dos Reis","sequence":"additional","affiliation":[{"name":"Department of Pathology, Barretos Cancer Hospital, Barretos 14784400, Brazil"}]},{"given":"Marcus","family":"Matsushita","sequence":"additional","affiliation":[{"name":"Department of Pathology, Barretos Cancer Hospital, Barretos 14784400, Brazil"}]},{"given":"Florinda","family":"Santos","sequence":"additional","affiliation":[{"name":"Department of Medical Oncology, Barretos Cancer Hospital, Barretos 14784400, Brazil"}]},{"given":"Denise Peixoto","family":"Guimar\u00e3es","sequence":"additional","affiliation":[{"name":"Molecular Oncology Research Center, Barretos Cancer Hospital, Barretos 14784400, Brazil"},{"name":"Department of Prevention, Barretos Cancer Hospital, Barretos 14784400, Brazil"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9639-7940","authenticated-orcid":false,"given":"Rui Manuel","family":"Reis","sequence":"additional","affiliation":[{"name":"Molecular Oncology Research Center, Barretos Cancer Hospital, Barretos 14784400, Brazil"},{"name":"Life and Health Sciences Research Institute (ICVS), School of Medicine, University of Minho, 4704553 Braga, Portugal"},{"name":"ICVS\/3B\u2019s\u2014PT Government Associate Laboratory, 4704553 Braga, Portugal"}]}],"member":"1968","published-online":{"date-parts":[[2022,1,5]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"209","DOI":"10.3322\/caac.21660","article-title":"Global Cancer Statistics 2020: GLOBOCAN Estimates of Incidence and Mortality Worldwide for 36 Cancers in 185 Countries","volume":"71","author":"Sung","year":"2021","journal-title":"Cancer J. 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