{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,17]],"date-time":"2026-06-17T22:56:52Z","timestamp":1781737012867,"version":"3.54.5"},"reference-count":25,"publisher":"MDPI AG","issue":"24","license":[{"start":{"date-parts":[[2025,12,7]],"date-time":"2025-12-07T00:00:00Z","timestamp":1765065600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["JCM"],"abstract":"<jats:p>Objectives: Pancreatic cystic lesions (PCLs) are increasingly detected due to the widespread use of imaging techniques. The identification of pancreatic mucinous cysts is especially important since these carry a risk of malignant transformation and require follow-up or surgical resection. The aim of this study was to determine the diagnostic yield of the molecular analysis of K-RAS (Kirsten RAt Sarcoma virus) and GNAS (Guanine Nucleotide-binding protein, Alpha Stimulating protein activity) gene mutations in pancreatic cyst fluid (PCF) obtained by endoscopic ultrasound (EUS)-guided fine needle aspiration (FNA). Methods: In this prospective trial, we assessed the sensitivity, specificity, and positive and negative predictive values of K-RAS and GNAS mutation analysis in differentiating mucinous versus non-mucinous cysts and the subsequent impact on decision-making in daily clinical practice. The reference standard used comprised the combination of morphology on cross-sectional imaging and EUS, string sign, cyst fluid cytology, intracystic carcinoembryonic antigen (CEA), and glucose levels, with subsequent correlation of surgical pathology in resected cases. Fluid samples of 47 cysts obtained by EUS-FNA over a 39-month period were analyzed. Mutation analysis of KRAS (exon 2) was performed in all cases, and additionally, GNAS (exon 8) in 28 cases using Sanger sequencing. Results: 33 out of 47 PCLs were classified as mucinous cysts and 14 as non-mucinous cysts defined using conventional standards, including morphological characteristics, string-sign, cytology, cyst fluid testing, and histology in resected cases. Of these 33 mucinous cysts, KRAS mutation was detected in 14 samples. A further 23 mucinous lesions were additionally tested for GNAS mutation, which was detected in 10 of the 23 cysts. A 42.4% sensitivity for KRAS and 43.5% for GNAS mutation analysis was calculated, with a specificity of 92.9% and 100%, respectively, for detecting mucinous lesions. The clinical management was altered through the genetic testing results in one single case. Conclusions: In this cohort, K-RAS and GNAS mutational analysis in cyst fluid did not improve the detection of mucinous pancreatic cysts significantly after conventional testing. However, the method may be useful due to its high specificity in uncertain cases.<\/jats:p>","DOI":"10.3390\/jcm14248671","type":"journal-article","created":{"date-parts":[[2025,12,8]],"date-time":"2025-12-08T16:20:29Z","timestamp":1765210829000},"page":"8671","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Clinical Benefits of KRAS\/GNAS Gene Mutation Analysis in Addition to Morphology and Conventional Cyst Fluid Testing in Differentiating Pancreatic Cysts"],"prefix":"10.3390","volume":"14","author":[{"given":"Gy\u00f6rgy","family":"Gyimesi","sequence":"first","affiliation":[{"name":"School of Doctoral Studies, Semmelweis University, 1082 Budapest, Hungary"},{"name":"Department of Gastroenterology, Spital Thurgau AG, 8596 M\u00fcnsterlingen, Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"B\u00e1nk","family":"Keczer","sequence":"additional","affiliation":[{"name":"Department of Surgery, Transplantation and Gastroenterology, Semmelweis University, 1082 Budapest, Hungary"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0009-1182-7242","authenticated-orcid":false,"given":"P\u00e9ter","family":"Rein","sequence":"additional","affiliation":[{"name":"Department of Surgery, Transplantation and Gastroenterology, Semmelweis University, 1082 Budapest, Hungary"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mikl\u00f3s","family":"Horv\u00e1th","sequence":"additional","affiliation":[{"name":"Department of Surgery, Transplantation and Gastroenterology, Division of Interventional Gastroenterology, Semmelweis University, 1082 Budapest, Hungary"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0002-7896-1541","authenticated-orcid":false,"given":"B\u00e1lint","family":"Gell\u00e9rt","sequence":"additional","affiliation":[{"name":"Department of Surgery, Transplantation and Gastroenterology, Division of Interventional Gastroenterology, Semmelweis University, 1082 Budapest, Hungary"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tam\u00e1s","family":"Marjai","sequence":"additional","affiliation":[{"name":"Department of Surgery, Transplantation and Gastroenterology, Semmelweis University, 1082 Budapest, Hungary"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0000-1370-3264","authenticated-orcid":false,"given":"Enik\u0151","family":"T\u00f3th","sequence":"additional","affiliation":[{"name":"Department of Surgery, Transplantation and Gastroenterology, Semmelweis University, 1082 Budapest, Hungary"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"\u00c1kos","family":"Sz\u0171cs","sequence":"additional","affiliation":[{"name":"Department of Surgery, Transplantation and Gastroenterology, Semmelweis University, 1082 Budapest, Hungary"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Attila","family":"Szij\u00e1rt\u00f3","sequence":"additional","affiliation":[{"name":"Department of Surgery, Transplantation and Gastroenterology, Semmelweis University, 1082 Budapest, Hungary"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tam\u00e1s","family":"Barbai","sequence":"additional","affiliation":[{"name":"Department of Pathology, Forensic and Insurance Medicine, Semmelweis University, 1082 Budapest, Hungary"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eszter","family":"Sz\u00e9kely","sequence":"additional","affiliation":[{"name":"Department of Pathology, Forensic and Insurance Medicine, Semmelweis University, 1082 Budapest, Hungary"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Istv\u00e1n","family":"Hritz","sequence":"additional","affiliation":[{"name":"Department of Surgery, Transplantation and Gastroenterology, Division of Interventional Gastroenterology, Semmelweis University, 1082 Budapest, Hungary"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,12,7]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"802","DOI":"10.2214\/AJR.07.3340","article-title":"Prevalence of unsuspected pancreatic cysts on MDCT","volume":"191","author":"Laffa","year":"2008","journal-title":"Am. J. Roentgenol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"2079","DOI":"10.1038\/ajg.2010.122","article-title":"Prevalence of incidental pancreatic cysts in the adult population on MR Imaging","volume":"105","author":"Lee","year":"2010","journal-title":"Am J. Gastroenterol."},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"De Oliviera, P.B., Puchnick, A., Szejnfeld, J., and Goldman, S.M. (2015). Prevalence of incidental pancreatic cysts on 3 tesla magnetic resonance [serial online]. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0121317"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"5700","DOI":"10.3748\/wjg.v27.i34.5700","article-title":"Updates in diagnosis and management of pancreatic cysts","volume":"27","author":"Lee","year":"2021","journal-title":"World J. Gastroenterol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"824","DOI":"10.1053\/j.gastro.2015.01.014","article-title":"American gastroenterological association technical review on the diagnosis and management of asymptomatic neoplastic pancreatic cysts","volume":"148","author":"Scheiman","year":"2015","journal-title":"Gastroenterology"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"e93","DOI":"10.1016\/j.suronc.2010.12.003","article-title":"Primary pancreatic cystic neoplasms revisited: Part II. Mucinous cystic neoplasms","volume":"20","author":"Sakorafas","year":"2011","journal-title":"Surg. Oncol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"915","DOI":"10.1177\/000313481408001001","article-title":"Mucinous cystic neoplasms of the pancreas: Are we overestimating malignant potential?","volume":"80","author":"Nguyen","year":"2014","journal-title":"Am. Surg."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"738","DOI":"10.1016\/j.pan.2017.07.007","article-title":"Revisions of international consensus Fukuoka guidelines for the management of IPMN of the pancreas","volume":"17","author":"Tanaka","year":"2017","journal-title":"Pancreatology"},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"(2018). The European Study Group on Cystic Tumors of the Pancreas: European evidence-based guidelines on pancreatic cystic neoplasms. Gut, 67, 789\u2013804.","DOI":"10.1136\/gutjnl-2018-316027"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"464","DOI":"10.1038\/ajg.2018.14","article-title":"ACG Clinical Guideline: Diagnosis and Management of Pancreatic Cysts","volume":"113","author":"Elta","year":"2018","journal-title":"Am. J. Gastroenterol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1330","DOI":"10.1053\/j.gastro.2004.02.013","article-title":"Diagnosis of pancreatic cystic neoplasms: A report of the cooperative pancreatic cyst study","volume":"126","author":"Brugge","year":"2004","journal-title":"Gastroenterology"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"626","DOI":"10.1055\/s-0034-1391484","article-title":"The string sign for diagnosis of mucinous pancreatic cysts","volume":"47","author":"Bick","year":"2015","journal-title":"Endoscopy"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"243","DOI":"10.14309\/ajg.0000000000002088","article-title":"Comparative performance of endoscopic ultrasound-based techniques in patients with pancreatic cystic lesions: A network meta-analysis","volume":"118","author":"Li","year":"2023","journal-title":"Am. J. Gastroenterol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"967","DOI":"10.1016\/S1542-3565(05)00409-X","article-title":"The role of pancreatic cyst fluid molecular analysis in predicting cyst pathology","volume":"3","author":"Khalid","year":"2005","journal-title":"Clin. Gastroenterol. Hepatol."},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Gillson, J., Ramaswamy, Y., Singh, G., Gorfe, A.A., Pavlakis, N., Samra, J., Mittal, A., and Sahni, S. (2020). Small Molecule KRAS Inhibitors: The Future for Targeted Pancreatic Cancer Therapy?. Cancers, 12.","DOI":"10.3390\/cancers12051341"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"692","DOI":"10.1038\/340692a0","article-title":"GTPase inhibiting mutations activate the alpha chain of Gs and stimulate adenylyl cyclase in human pituitary tumours","volume":"340","author":"Landis","year":"1989","journal-title":"Nature"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"5459","DOI":"10.1210\/en.2004-0865","article-title":"Minireview: GNAS: Normal and abnormal functions","volume":"145","author":"Weinstein","year":"2004","journal-title":"Endocrinology"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1019","DOI":"10.1016\/j.gie.2020.12.014","article-title":"Molecular analysis of EUS-acquired pancreatic cyst fluid for KRAS and GNAS mutations for diagnosis of intraductal papillary mucinous neoplasia and mucinous cystic lesions: A systematic review and meta-analysis","volume":"93","author":"McCarty","year":"2021","journal-title":"Gastrointest. Endosc."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"183","DOI":"10.1016\/j.pan.2012.04.004","article-title":"International Association of Pancreatology. International consensus guidelines 2012 for the management of IPMN and MCN of the pancreas","volume":"12","author":"Tanaka","year":"2012","journal-title":"Pancreatology"},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"American Gastroenterological Association, and AGA Section (2015). Managing Pancreatic Cysts: A Patient Guide. Gastroenterology, 149, 498\u2013499.","DOI":"10.1053\/j.gastro.2015.06.042"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1611881","DOI":"10.3389\/pore.2024.1611881","article-title":"Diagnostic performance of intracystic carcinoembryonic antigen (CEA) versus glucose in differentiation of mucinous and non-mucinous pancreatic cysts","volume":"30","author":"Gyimesi","year":"2024","journal-title":"Pathol. Oncol. Res."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"909","DOI":"10.1038\/ajg.2015.148","article-title":"Cyst Fluid Glucose is Rapidly Feasible and Accurate in Diagnosing Mucinous Pancreatic Cysts","volume":"110","author":"Zikos","year":"2015","journal-title":"Am. J. Gastroenterol."},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Carmicheal, J., Patel, A., Dalal, V., Atri, P., Dhaliwal, A.S., Wittel, U.A., Malafa, M.P., Talmon, G., Swanson, B.J., and Singh, S. (2020). Elevating pancreatic cystic lesion stratification: Current and future pancreatic cancer biomarker(s). Biochim. Biophys. Acta Rev. Cancer., 1873.","DOI":"10.1016\/j.bbcan.2019.188318"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"2351","DOI":"10.1007\/s10620-018-5128-y","article-title":"Clinical Impact of KRAS and GNAS Analysis Added to CEA and Cytology in Pancreatic Cystic Fluid Obtained by EUS-FNA","volume":"63","author":"Faias","year":"2018","journal-title":"Dig. Dis. Sci."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"56","DOI":"10.4253\/wjge.v13.i2.56","article-title":"Molecular analysis of pancreatic cystic neoplasm in routine clinical practice","volume":"13","year":"2021","journal-title":"World J. Gastrointest. Endosc."}],"container-title":["Journal of Clinical Medicine"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2077-0383\/14\/24\/8671\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,12,8]],"date-time":"2025-12-08T16:50:24Z","timestamp":1765212624000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2077-0383\/14\/24\/8671"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,12,7]]},"references-count":25,"journal-issue":{"issue":"24","published-online":{"date-parts":[[2025,12]]}},"alternative-id":["jcm14248671"],"URL":"https:\/\/doi.org\/10.3390\/jcm14248671","relation":{},"ISSN":["2077-0383"],"issn-type":[{"value":"2077-0383","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,12,7]]}}}