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The RNA-based Lymph2Cx assay has been validated as a reliable surrogate of high-throughput gene expression profiling assays for distinguishing between GCB and ABC DLBCL and provides reliable results from formalin-fixed, paraffin-embedded (FFPE) material. This test has been previously used in clinical trials, but experience from real-world routine application is rare. We routinely applied the Lymph2Cx assay to day-to-day diagnostics on a series of 147 aggressive B-cell lymphoma cases and correlated our results with the immunohistochemical subclassification using the Hans algorithm and fluorescence in situ hybridization findings using break-apart probes for\n                    <jats:italic>MYC<\/jats:italic>\n                    ,\n                    <jats:italic>BCL2<\/jats:italic>\n                    , and\n                    <jats:italic>BCL6<\/jats:italic>\n                    . The routine use of the Lymph2Cx assay had a high technical success rate (94.6%) with a low rate of failure due to poor material and\/or RNA quality. The Lymph2Cx assay was discordant with the Hans algorithm in 18% (23 of 128 cases). Discordant cases were mainly classified as GCB by the Hans algorithm and as ABC by Lymph2Cx (n\u2009=\u200911, 8.6%). Only 5 cases (3.9%) were classified as non-GCB by the Hans algorithm and as GCB by Lymph2Cx. Additionally, 5.5% of cases (n\u2009=\u20097) were left unclassified by Lymph2Cx, whereas they were defined as GCB (n\u2009=\u20094) or non-GCB (n\u2009=\u20093) by the Hans algorithm. Our data support the routine applicability of the Lymph2Cx assay.\n                  <\/jats:p>","DOI":"10.1007\/s00428-022-03420-6","type":"journal-article","created":{"date-parts":[[2022,10,11]],"date-time":"2022-10-11T09:03:36Z","timestamp":1665479016000},"page":"935-943","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":13,"title":["Routine application of the Lymph2Cx assay for the subclassification of aggressive B-cell lymphoma: report of a prospective real-world series"],"prefix":"10.1007","volume":"481","author":[{"given":"Alberto","family":"Zam\u00f2","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elena","family":"Gerhard-Hartmann","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"German","family":"Ott","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ioannis","family":"Anagnostopoulos","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"David W.","family":"Scott","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andreas","family":"Rosenwald","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5535-2412","authenticated-orcid":false,"given":"Hilka","family":"Rauert-Wunderlich","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2022,10,11]]},"reference":[{"key":"3420_CR1","doi-asserted-by":"publisher","first-page":"1361","DOI":"10.1182\/blood.V84.5.1361.1361","volume":"84","author":"NL Harris","year":"1994","unstructured":"Harris NL, Jaffe ES, Stein H, Banks PM, Chan JK, Cleary ML et al (1994) A revised European-American classification of lymphoid neoplasms: a proposal from the International Lymphoma Study Group. 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