{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,27]],"date-time":"2025-10-27T17:42:55Z","timestamp":1761586975422,"version":"build-2065373602"},"reference-count":41,"publisher":"Public Library of Science (PLoS)","issue":"10","license":[{"start":{"date-parts":[[2025,10,27]],"date-time":"2025-10-27T00:00:00Z","timestamp":1761523200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS One"],"abstract":"<jats:sec id=\"sec001\">\n                    <jats:title>Background<\/jats:title>\n                    <jats:p>Cholangiocarcinoma (CCA) is a deadly cancer often detected late. Current diagnostic methods, such as ultrasound and invasive biopsies, have limitations; there is a critical need for a rapid, minimally invasive and effective strategy for the early diagnosis and staging of CCA.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec id=\"sec002\">\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      We aimed to address this need using serum samples and label-free surface-enhanced Raman spectroscopy (SERS) combined with machine learning. CCA development was induced in hamsters using a combination of\n                      <jats:italic>Opisthorchis viverrini<\/jats:italic>\n                      infection and administration of\n                      <jats:italic>N<\/jats:italic>\n                      -nitrosodimethylamine, with induction time courses spanning 1\u20135 month(s). Normal and pathological stages (inflammation, precancerous lesion, and CCA) were assigned based on histopathological features, as well as the expression of cytokeratin 19 and alpha-fetoprotein. Raman spectra were subjected to dimensionality reduction using principal component analysis, and diagnostic clusters were acquired using partial least-squares discriminant analysis.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec id=\"sec003\">\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>Histopathological analysis confirmed a clear path towards CCA, initiated by marked inflammation, progressing to include significant cholangiofibrosis and cholangiofibroma in the precancerous stage, and culminating in definitive CCA tumor development. The integration of SERS and machine learning achieved a diagnostic sensitivity of 93%, specificity of 95%, and accuracy of \u2265 67% for precancerous lesions and CCA, with an area under the receiver operating characteristic curve exceeding 0.67.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec id=\"sec004\">\n                    <jats:title>Conclusions<\/jats:title>\n                    <jats:p>Our findings demonstrate that this cost-effective, label-free SERS approach can accurately detect precancerous and cancerous stages of cholangiocarcinoma in a hamster model, highlighting its strong potential for future development as a community-based screening tool.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1371\/journal.pone.0334916","type":"journal-article","created":{"date-parts":[[2025,10,27]],"date-time":"2025-10-27T17:40:48Z","timestamp":1761586848000},"page":"e0334916","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":0,"title":["Minimally invasive detection of early-stage opisthorchiasis-associated cholangiocarcinoma using label-free surface-enhanced Raman spectroscopy (SERS) of hamster serum"],"prefix":"10.1371","volume":"20","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-6495-0951","authenticated-orcid":true,"given":"Apisit","family":"Chaidee","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Suppakrit","family":"Kongsintaweesuk","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Thatsanapong","family":"Pongking","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Keerapach","family":"Tunbenjasiri","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Aye Myat","family":"Mon","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Chawalit","family":"Pairojkul","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0001-9735-028X","authenticated-orcid":true,"given":"Pakornkiat","family":"Tanasuka","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tullayakorn","family":"Plengsuriyakarn","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kesara","family":"Na-Bangchang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Naruechar","family":"Charoenram","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"David","family":"Blair","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8187-7247","authenticated-orcid":true,"given":"Somchai","family":"Pinlaor","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"340","published-online":{"date-parts":[[2025,10,27]]},"reference":[{"issue":"6","key":"pone.0334916.ref001","doi-asserted-by":"crossref","first-page":"1215","DOI":"10.1053\/j.gastro.2013.10.013","article-title":"Pathogenesis, diagnosis, and management of cholangiocarcinoma","volume":"145","author":"SI Ilyas","year":"2013","journal-title":"Gastroenterology"},{"issue":"5","key":"pone.0334916.ref002","doi-asserted-by":"crossref","first-page":"594","DOI":"10.1634\/theoncologist.2015-0446","article-title":"Forty-Year Trends in Cholangiocarcinoma Incidence in the U.S.: Intrahepatic Disease on the Rise","volume":"21","author":"SK Saha","year":"2016","journal-title":"Oncologist"},{"issue":"9","key":"pone.0334916.ref003","doi-asserted-by":"crossref","first-page":"557","DOI":"10.1038\/s41575-020-0310-z","article-title":"Cholangiocarcinoma 2020: the next horizon in mechanisms and management","volume":"17","author":"JM Banales","year":"2020","journal-title":"Nat Rev Gastroenterol Hepatol"},{"issue":"5","key":"pone.0334916.ref004","doi-asserted-by":"crossref","first-page":"316","DOI":"10.1002\/jhbp.64","article-title":"Histological confirmation of periductal fibrosis from ultrasound diagnosis in cholangiocarcinoma patients","volume":"21","author":"N Chamadol","year":"2014","journal-title":"J Hepatobiliary Pancreat Sci"},{"issue":"3","key":"pone.0334916.ref005","doi-asserted-by":"crossref","first-page":"579","DOI":"10.1111\/j.1349-7006.2009.01458.x","article-title":"Epidemiology of cholangiocarcinoma: an update focusing on risk factors","volume":"101","author":"H-R Shin","year":"2010","journal-title":"Cancer Sci"},{"issue":"3","key":"pone.0334916.ref006","doi-asserted-by":"crossref","first-page":"540","DOI":"10.1128\/CMR.17.3.540-552.2004","article-title":"Clonorchiasis and cholangiocarcinoma: etiologic relationship and imaging diagnosis","volume":"17","author":"BI Choi","year":"2004","journal-title":"Clin Microbiol Rev"},{"issue":"1","key":"pone.0334916.ref007","doi-asserted-by":"crossref","first-page":"3","DOI":"10.3892\/ol.2014.2696","article-title":"Diagnostic and prognostic serum marker of cholangiocarcinoma (Review)","volume":"9","author":"X Zeng","year":"2015","journal-title":"Oncol Lett"},{"issue":"23","key":"pone.0334916.ref008","doi-asserted-by":"crossref","first-page":"15361","DOI":"10.1021\/acs.analchem.0c02656","article-title":"Monitoring the Progression of Liver Fluke-Induced Cholangiocarcinoma in a Hamster Model Using Synchrotron FTIR Microspectroscopy and Focal Plane Array Infrared Imaging","volume":"92","author":"P Chatchawal","year":"2020","journal-title":"Anal Chem"},{"key":"pone.0334916.ref009","first-page":"8619342","article-title":"Raman Spectroscopy and Imaging for Cancer Diagnosis","volume":"2018","author":"S Cui","year":"2018","journal-title":"J Healthc Eng"},{"issue":"4","key":"pone.0334916.ref010","doi-asserted-by":"crossref","first-page":"708","DOI":"10.1016\/j.cbpa.2013.05.021","article-title":"Molecular imaging of live cells by Raman microscopy","volume":"17","author":"AF Palonpon","year":"2013","journal-title":"Curr Opin Chem Biol"},{"issue":"10","key":"pone.0334916.ref011","doi-asserted-by":"crossref","DOI":"10.1371\/journal.pone.0275362","article-title":"Rapid label-free detection of cholangiocarcinoma from human serum using Raman spectroscopy","volume":"17","author":"P Suksuratin","year":"2022","journal-title":"PLoS One"},{"key":"pone.0334916.ref012","doi-asserted-by":"crossref","first-page":"123702","DOI":"10.1016\/j.saa.2023.123702","article-title":"Application of surface-enhanced Raman spectroscopy to human serum for diagnosing liver cancer","volume":"308","author":"Q Ou","year":"2024","journal-title":"Spectrochim Acta A Mol Biomol Spectrosc"},{"key":"pone.0334916.ref013","doi-asserted-by":"crossref","first-page":"124046","DOI":"10.1016\/j.saa.2024.124046","article-title":"Surface-enhanced Raman spectroscopy of the filtrate portions of the blood serum samples of breast cancer patients obtained by using 30 kDa filtration device","volume":"311","author":"H Hajab","year":"2024","journal-title":"Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy"},{"issue":"4","key":"pone.0334916.ref014","doi-asserted-by":"crossref","first-page":"2020","DOI":"10.1021\/acssensors.3c02810","article-title":"Optical Nanobiosensor Based on Surface-Enhanced Raman Spectroscopy and Catalytic Hairpin Assembly for Early-Stage Lung Cancer Detection via Blood Circular RNA","volume":"9","author":"L Xu","year":"2024","journal-title":"ACS Sens"},{"issue":"27","key":"pone.0334916.ref015","doi-asserted-by":"crossref","first-page":"11593","DOI":"10.1039\/D1NR00708D","article-title":"Surface-enhanced Raman spectroscopy for bioanalysis and diagnosis","volume":"13","author":"MA Tahir","year":"2021","journal-title":"Nanoscale"},{"key":"pone.0334916.ref016","first-page":"248","article-title":"SERS sensing for cancer biomarker: Approaches and directions","volume":"34","author":"L V\u00e1zquez-Iglesias","year":"2023","journal-title":"Bioact Mater"},{"issue":"9","key":"pone.0334916.ref017","doi-asserted-by":"crossref","first-page":"12276","DOI":"10.1021\/acsanm.2c02392","article-title":"Label-Free Sensing with Metal Nanostructure-Based Surface-Enhanced Raman Spectroscopy for Cancer Diagnosis","volume":"5","author":"M Constantinou","year":"2022","journal-title":"ACS Appl Nano Mater"},{"issue":"7","key":"pone.0334916.ref018","doi-asserted-by":"crossref","first-page":"1577","DOI":"10.1111\/j.1476-5381.2010.00872.x","article-title":"Animal research: reporting in vivo experiments: the ARRIVE guidelines","volume":"160","author":"C Kilkenny","year":"2010","journal-title":"Br J Pharmacol"},{"issue":"1","key":"pone.0334916.ref019","doi-asserted-by":"crossref","DOI":"10.1186\/s12645-023-00155-0","article-title":"Curcumin-loaded nanocomplexes alleviate the progression of fluke-related cholangiocarcinoma in hamsters","volume":"14","author":"C Jantawong","year":"2023","journal-title":"Cancer Nano"},{"key":"pone.0334916.ref020","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1038\/s41536-017-0018-z","article-title":"Understanding liver regeneration to bring new insights to the mechanisms driving cholangiocarcinoma","volume":"2","author":"RV Guest","year":"2017","journal-title":"NPJ Regen Med"},{"issue":"4","key":"pone.0334916.ref021","doi-asserted-by":"crossref","first-page":"339","DOI":"10.1293\/tox.2021-0032","article-title":"Spontaneous cholangiofibrosis adjacent to a dilated common bile duct with intestinal metaplasia in a Royal College of Surgeons rat","volume":"34","author":"K Ozaki","year":"2021","journal-title":"J Toxicol Pathol"},{"issue":"5","key":"pone.0334916.ref022","doi-asserted-by":"crossref","first-page":"101","DOI":"10.1002\/cld.345","article-title":"Biomarkers in cholangiocarcinoma","volume":"3","author":"JA Marrero","year":"2014","journal-title":"Clin Liver Dis (Hoboken)"},{"issue":"2","key":"pone.0334916.ref023","doi-asserted-by":"crossref","first-page":"128","DOI":"10.1007\/s11901-015-0261-y","article-title":"Screening for hepatocellular carcinoma and cholangiocarcinoma: Can biomarkers replace imaging?","volume":"14","author":"ME Lozada","year":"2015","journal-title":"Curr Hepatol Rep"},{"key":"pone.0334916.ref024","doi-asserted-by":"crossref","DOI":"10.1177\/0192623310386499","article-title":"Proliferative and nonproliferative lesions of the rat and mouse hepatobiliary system","volume":"38","author":"B Thoolen","year":"2010","journal-title":"Toxicol Pathol"},{"key":"pone.0334916.ref025","doi-asserted-by":"crossref","first-page":"116063","DOI":"10.1016\/j.bios.2024.116063","article-title":"Determination of latent tuberculosis infection from plasma samples via label-free SERS sensors and machine learning","volume":"250","author":"P Eiamchai","year":"2024","journal-title":"Biosens Bioelectron"},{"issue":"11","key":"pone.0334916.ref026","doi-asserted-by":"crossref","first-page":"11296","DOI":"10.1021\/acsomega.4c11078","article-title":"Highly Accurate and Robust Early Stage Detection of Cholangiocarcinoma Using Near-Lossless SERS Signal Processing with Machine Learning and 2D CNN for Point-of-care Mobile Application","volume":"10","author":"P Danvirutai","year":"2025","journal-title":"ACS Omega"},{"issue":"15","key":"pone.0334916.ref027","doi-asserted-by":"crossref","first-page":"4789","DOI":"10.1140\/epjs\/s11734-024-01449-z","article-title":"Deep learning of surface-enhanced Raman spectroscopy data for multiple sclerosis diagnostics","volume":"234","author":"AV Zakharov","year":"2025","journal-title":"Eur Phys J Spec Top"},{"issue":"17","key":"pone.0334916.ref028","doi-asserted-by":"crossref","first-page":"15385","DOI":"10.1021\/acsanm.3c01442","article-title":"Using Machine Learning and Silver Nanoparticle-Based Surface-Enhanced Raman Spectroscopy for Classification of Cardiovascular Disease Biomarkers","volume":"6","author":"K Dixon","year":"2023","journal-title":"ACS Appl Nano Mater"},{"issue":"2","key":"pone.0334916.ref029","doi-asserted-by":"crossref","first-page":"233","DOI":"10.3390\/diagnostics13020233","article-title":"Diagnosis of Cholangiocarcinoma","volume":"13","author":"DW Shin","year":"2023","journal-title":"Diagnostics (Basel)"},{"issue":"11","key":"pone.0334916.ref030","doi-asserted-by":"crossref","first-page":"2576","DOI":"10.1002\/ijc.25266","article-title":"Involvement of MMP-9 in peribiliary fibrosis and cholangiocarcinogenesis via Rac1-dependent DNA damage in a hamster model","volume":"127","author":"S Prakobwong","year":"2010","journal-title":"Int J Cancer"},{"issue":"1","key":"pone.0334916.ref031","doi-asserted-by":"crossref","first-page":"40","DOI":"10.1038\/s41575-020-0342-4","article-title":"Liver regeneration: biological and pathological mechanisms and implications","volume":"18","author":"GK Michalopoulos","year":"2021","journal-title":"Nat Rev Gastroenterol Hepatol"},{"issue":"1","key":"pone.0334916.ref032","doi-asserted-by":"crossref","first-page":"401","DOI":"10.7314\/APJCP.2016.17.1.401","article-title":"Cholangiocarcinoma Patient Outcome in Northeastern Thailand: Single-Center Prospective Study","volume":"17","author":"V Luvira","year":"2016","journal-title":"Asian Pac J Cancer Prev"},{"key":"pone.0334916.ref033","doi-asserted-by":"crossref","first-page":"459","DOI":"10.1186\/s12885-015-1475-7","article-title":"Cohort profile: cholangiocarcinoma screening and care program (CASCAP)","volume":"15","author":"N Khuntikeo","year":"2015","journal-title":"BMC Cancer"},{"issue":"4","key":"pone.0334916.ref034","doi-asserted-by":"crossref","first-page":"664","DOI":"10.1038\/nprot.2016.036","article-title":"Using Raman spectroscopy to characterize biological materials","volume":"11","author":"HJ Butler","year":"2016","journal-title":"Nat Protoc"},{"issue":"8","key":"pone.0334916.ref035","doi-asserted-by":"crossref","first-page":"1343","DOI":"10.3390\/s7081343","article-title":"Raman Spectroscopy Cell-based Biosensors","volume":"7","author":"I Notingher","year":"2007","journal-title":"Sensors"},{"issue":"22","key":"pone.0334916.ref036","doi-asserted-by":"crossref","first-page":"6721","DOI":"10.1039\/C9AN01772K","article-title":"Identification of early inflammatory changes in the tympanic membrane with Raman spectroscopy","volume":"144","author":"SP Singh","year":"2019","journal-title":"Analyst"},{"issue":"14","key":"pone.0334916.ref037","doi-asserted-by":"crossref","first-page":"1909","DOI":"10.3390\/cells12141909","article-title":"Early Detection of Pre-Cancerous and Cancerous Cells Using Raman Spectroscopy-Based Machine Learning","volume":"12","author":"U Sharaha","year":"2023","journal-title":"Cells"},{"issue":"8","key":"pone.0334916.ref038","doi-asserted-by":"crossref","first-page":"1125","DOI":"10.1038\/s41416-021-01659-5","article-title":"Raman spectroscopy: current applications in breast cancer diagnosis, challenges and future prospects","volume":"126","author":"K Hanna","year":"2022","journal-title":"Br J Cancer"},{"issue":"4","key":"pone.0334916.ref039","doi-asserted-by":"crossref","first-page":"691","DOI":"10.1007\/s10555-018-9770-9","article-title":"Applications of Raman spectroscopy in cancer diagnosis","volume":"37","author":"GW Auner","year":"2018","journal-title":"Cancer Metastasis Rev"},{"issue":"6","key":"pone.0334916.ref040","doi-asserted-by":"crossref","first-page":"4274","DOI":"10.1529\/biophysj.103.038604","article-title":"Raman spectroscopy detects biochemical changes due to proliferation in mammalian cell cultures","volume":"88","author":"KW Short","year":"2005","journal-title":"Biophys J"},{"issue":"9","key":"pone.0334916.ref041","doi-asserted-by":"crossref","first-page":"1855","DOI":"10.1007\/s10103-020-03218-5","article-title":"Raman spectroscopic study of cervical precancerous lesions and cervical cancer","volume":"36","author":"J Wang","year":"2021","journal-title":"Lasers Med Sci"}],"container-title":["PLOS One"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0334916","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,27]],"date-time":"2025-10-27T17:40:53Z","timestamp":1761586853000},"score":1,"resource":{"primary":{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0334916"}},"subtitle":[],"editor":[{"given":"Maria Y","family":"Pakharukova","sequence":"first","affiliation":[],"role":[{"role":"editor","vocabulary":"crossref"}]}],"short-title":[],"issued":{"date-parts":[[2025,10,27]]},"references-count":41,"journal-issue":{"issue":"10","published-online":{"date-parts":[[2025,10,27]]}},"URL":"https:\/\/doi.org\/10.1371\/journal.pone.0334916","relation":{},"ISSN":["1932-6203"],"issn-type":[{"value":"1932-6203","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,10,27]]}}}