{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,7]],"date-time":"2026-07-07T14:25:02Z","timestamp":1783434302543,"version":"3.54.6"},"reference-count":32,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2025,3,14]],"date-time":"2025-03-14T00:00:00Z","timestamp":1741910400000},"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>Background: Methods such as cytogenetics and immunocytology\/immunohistology provide essential diagnostic insights but may be limited in ambiguous cases of mature B-cell lymphoma. Next-generation sequencing (NGS) has emerged as a potential tool to improve diagnostics. Methods: We validated the analytical performance of a lymphoid customized NGS panel. Clinical validation was conducted in 226 patients with suspected mature B-cell lymphoma with potential bone marrow involvement across multiple clinically relevant scenarios. Results: NGS (1) achieved 100% sensitivity and specificity with high reproducibility (r = 0.995), confirming its analytical performance. (2) It reliably detected WHO-classified markers, including BRAF mutations in all hairy cell leukemia cases, MYD88\/CXCR4 mutations in lymphoplasmacytic lymphoma, and absence of BRAF mutations in splenic B-cell lymphoma with prominent nucleoli. (3) In lymphoma exclusion diagnostics, NGS identified mutations in previously undiagnosed cases, including a BCORL1 mutation leading to reclassification as marginal zone lymphoma. (4) Among 105 confirmed lymphomas, 65% harbored mutations, with detection rates highest in HCL and LPL (100%) and CLL (62%), while follicular lymphoma showed no detectable mutations. (5) In cases with non-interpretable cytogenetics, NGS detected pathogenic variants in 61% of patients, compensating for inconclusive findings. (6) In cases with limited morphological assessment, NGS identified relevant mutations in 70%, outperforming cytogenetics (30%; p = 0.0256, OR = 5.44). Conclusions: NGS enhances the diagnostic accuracy of mature B-cell lymphomas by complementing traditional methods, refining WHO-classified subtypes, and improving detection in cases with inconclusive cytogenetics or morphology. NGS may reduce the need for unnecessary bone marrow re-punctures by providing additional information in ambiguous cases.<\/jats:p>","DOI":"10.3390\/diagnostics15060727","type":"journal-article","created":{"date-parts":[[2025,3,14]],"date-time":"2025-03-14T07:02:16Z","timestamp":1741935736000},"page":"727","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Diagnostic Implications of NGS-Based Molecular Profiling in Mature B-Cell Lymphomas with Potential Bone Marrow Involvement"],"prefix":"10.3390","volume":"15","author":[{"given":"Bernhard","family":"Strasser","sequence":"first","affiliation":[{"name":"Institute of Clinical Chemistry and Laboratory Medicine, Klinikum Wels-Grieskirchen 1, 4600 Wels, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sebastian","family":"Mustafa","sequence":"additional","affiliation":[{"name":"Institute of Clinical Chemistry and Laboratory Medicine, Klinikum Wels-Grieskirchen 1, 4600 Wels, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Josef","family":"Seier","sequence":"additional","affiliation":[{"name":"Institute of Clinical Chemistry and Laboratory Medicine, Klinikum Wels-Grieskirchen 1, 4600 Wels, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Erich","family":"Wimmer","sequence":"additional","affiliation":[{"name":"Institute of Clinical Chemistry and Laboratory Medicine, Klinikum Wels-Grieskirchen 1, 4600 Wels, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Josef","family":"Tomasits","sequence":"additional","affiliation":[{"name":"Department of Internal Medicine, Johannes Kepler University, 4040 Linz, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,3,14]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1720","DOI":"10.1038\/s41375-022-01620-2","article-title":"The 5th edition of the World Health Organization Classification of Haematolymphoid Tumours: Lymphoid Neoplasms","volume":"36","author":"Alaggio","year":"2022","journal-title":"Leukemia"},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Rodenas Quinonero, I., Marco-Ayala, J., Chen-Liang, T.H., de la Cruz-Vicente, F., Baumann, T., Navarro, J.T., Martin Garcia-Sancho, A., Martin-Santos, T., Lopez-Jimenez, J., and Andreu, R. 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