The aim of the study was to compare the prognostic significance of lymph node status of patients with lung cancer analyzed by three different methods: hematoxylin and eosin (H&E), immunohistochemistry of cytokeratin 19 (IHC CK19), and One-Step Nucleic Acid Amplification (OSNA). The clinical relevance of the results was evaluated based on relation to prognosis; the disease-free interval (DFI) and overall survival (OS) were analyzed. During radical surgical treatment, a total of 1426 lymph nodes were obtained from 100 patients, creating 472 groups of nodes (4-5 groups per patient) and examined by H&E, IHC CK19 and OSNA. The median follow-up was 44 months. Concordant results on the lymph node status of the H&E, IHC CK19 and OSNA examinations were reported in 78% of patients. We recorded shorter OS in patients with positive results provided by both OSNA and H&E. The study demonstrated a higher percentage of detected micrometastases in lymph nodes by the OSNA method. However, the higher sensitivity of the OSNA, with the cut-off value 250 copies of mRNA of CK19/µL, resulted in a lower association of OSNA positivity with progress of the disease compared to H&E. Increasing the cut-off to 615 copies resulted in an increase in concordance between the OSNA and H&E, which means that the higher cut-off is more relevant in the case of lung tumors.

Biomedical Center Faculty of Medicine in Pilsen Charles University alej Svobody 76 323 00 Plzen Czech Republic

Department of Biology Faculty of Medicine in Pilsen Charles University alej Svobody 76 323 00 Plzen Czech Republic

Department of Immunochemistry Diagnostics Faculty Hospital Plzen Faculty of Medicine in Pilsen Charles University Edvarda Benese 13 305 99 Plzen Czech Republic

Department of Medical Chemistry and Biochemistry Faculty of Medicine in Pilsen Charles University Karlovarska 48 301 66 Plzen Czech Republic

Department of Pathology Faculty Hospital Plzen Faculty of Medicine in Pilsen Charles University Edvarda Benese 13 305 99 Plzen Czech Republic

Department of Surgery Faculty Hospital Plzen Faculty of Medicine in Pilsen Charles University alej Svobody 80 304 60 Plzen Czech Republic

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Croner R.S., Geppert C.-I., Bader F.G., Nitsche U., Späth C., Rosenberg R., Zettl A., Matias-Guiu X., Tarragona J., Güller U., et al. Molecular staging of lymph node-negative colon carcinomas by one-step nucleic acid amplification (OSNA) results in upstaging of a quarter of patients in a prospective, European, multicentre study. Br. J. Cancer. 2014;110:2544–2550. doi: 10.1038/bjc.2014.170. PubMed DOI PMC

Osako T., Iwase T., Kimura K., Horii R., Akiyama F. Sentinel node tumour burden quantified based on cytokeratin 19 mRNA copy number predicts non-sentinel node metastases in breast cancer: Molecular whole-node analysis of all removed nodes. Eur. J. Cancer. 2013;49:1187–1195. doi: 10.1016/j.ejca.2012.11.022. PubMed DOI

Hayama M., Chida M., Karube Y., Tamura M., Kobayashi S., Oyaizu T., Honma K. One-step nucleic acid amplification for detection of lymph node metastasis in lung cancer. Ann. Thorac. Cardiovasc. Surg. 2014;20:181–184. doi: 10.5761/atcs.oa.12.02224. PubMed DOI

Vogelaar F.J., Reimers M.S., van der Linden R.L.A., van der Linden J.C., Smit V.T.H.B.M., Lips D.J., van de Velde C.J.H., Bosscha K. The Diagnostic Value of One-Step Nucleic acid Amplification (OSNA) for Sentinel Lymph Nodes in Colon Cancer Patients. Ann. Surg. Oncol. 2014;21:3924–3930. doi: 10.1245/s10434-014-3820-5. PubMed DOI

Chaudhry A., Williams S., Cook J., Jenkins M., Sohail M., Calder C., Winters Z.E., Rayter Z. The real-time intra-operative evaluation of sentinel lymph nodes in breast cancer patients using One Step Nucleic Acid Amplification (OSNA) and implications for clinical decision-making. Eur. J. Surg. Oncol. 2014;40:150–157. doi: 10.1016/j.ejso.2013.12.007. PubMed DOI

Le Frère-Belda M.-A., Bats A.-S., Gillaizeau F., Poulet B., Clough K.B., Nos C., Peoc’h M., Seffert P., Bouteille C., Leroux A., et al. Diagnostic performance of one-step nucleic acid amplification for intraoperative sentinel node metastasis detection in breast cancer patients. Int. J. Cancer. 2012;130:2377–2386. doi: 10.1002/ijc.26291. PubMed DOI

Güller U., Zettl A., Worni M., Langer I., Cabalzar-Wondberg D., Viehl C.T., Demartines N., Zuber M. Molecular investigation of lymph nodes in colon cancer patients using one-step nucleic acid amplification (OSNA): A new road to better staging? Cancer. 2012;118:6039–6045. doi: 10.1002/cncr.27667. PubMed DOI PMC

Osako T., Iwase T., Kimura K., Masumura K., Horii R., Akiyama F. Incidence and possible pathogenesis of sentinel node micrometastases in ductal carcinoma in situ of the breast detected using molecular whole lymph node assay. Br. J. Cancer. 2012;106:1675–1681. doi: 10.1038/bjc.2012.168. PubMed DOI PMC

Remoundos D.D., Ng V.V., Wilson H.A., Ahmed F., Chia Y., Cunnick G.H. The use of one step nucleic-acid amplification (OSNA) in clinical practice: A single-centre study. Breast. 2013;22:162–167. doi: 10.1016/j.breast.2012.11.003. PubMed DOI

Yamamoto N., Daito M., Hiyama K., Ding J., Nakabayashi K., Otomo Y., Tsujimoto M., Matsuura N., Kato Y. An Optimal mRNA Marker for OSNA (One-step Nucleic Acid Amplification) Based Lymph Node Metastasis Detection in Colorectal Cancer Patients. Jpn. J. Clin. Oncol. 2013;43:264–270. doi: 10.1093/jjco/hys227. PubMed DOI

Kosťun J., Pešta M., Sláma J., Slunéčko R., Vlasák P., Bouda J., Novotný Z., Topolčan O., Kučera R., Kulda V., et al. One-step nucleic acid amplification vs ultrastaging in the detection of sentinel lymph node metastasis in endometrial cancer patients. J. Surg. Oncol. 2019;119:361–369. doi: 10.1002/jso.25322. PubMed DOI

Gęca K., Rawicz-Pruszyński K., Mielko J., Mlak R., Sędłak K., Polkowski W.P. Rapid Detection of Free Cancer Cells in Intraoperative Peritoneal Lavage Using One-Step Nucleic Acid Amplification (OSNA) in Gastric Cancer Patients. Cells. 2020;9:2168. doi: 10.3390/cells9102168. PubMed DOI PMC

Del Carmen S., Gatius S., Franch-Arcas G., Baena J.A., Gonzalez O., Zafon C., Cuevas D., Valls J., Pérez A., Martinez M., et al. Concordance study between one-step nucleic acid amplification and morphologic techniques to detect lymph node metastasis in papillary carcinoma of the thyroid. Hum. Pathol. 2016;48:132–141. doi: 10.1016/j.humpath.2015.09.020. PubMed DOI

Medas F., Coni P., Podda F., Salaris C., Cappellacci F., Faa G., Calò P.G. Evaluation of accuracy of one-step nucleic acid amplification (OSNA) in diagnosis of lymph node metastases of papillary thyroid carcinoma. Diagnostic study. Ann. Med. Surg. 2012. 2019;46:17–22. doi: 10.1016/j.amsu.2019.08.006. PubMed DOI PMC

Inoue M., Hiyama K., Nakabayashi K., Morii E., Minami M., Sawabata N., Shintani Y., Nakagiri T., Susaki Y., Maeda J., et al. An accurate and rapid detection of lymph node metastasis in non-small cell lung cancer patients based on one-step nucleic acid amplification assay. Lung Cancer. 2012;78:212–218. doi: 10.1016/j.lungcan.2012.08.018. PubMed DOI

Masai K., Nakagawa K., Yoshida A., Sakurai H., Watanabe S., Asamura H., Tsuta K. Cytokeratin 19 expression in primary thoracic tumors and lymph node metastases. Lung Cancer. 2014;86:318–323. doi: 10.1016/j.lungcan.2014.09.018. PubMed DOI

Nakagawa K., Asamura H., Tsuta K., Nagai K., Yamada E., Ishii G., Mitsudomi T., Ito A., Higashiyama M., Tomita Y., et al. The novel one-step nucleic acid amplification (OSNA) assay for the diagnosis of lymph node metastasis in patients with non-small cell lung cancer (NSCLC): Results of a multicenter prospective study. Lung Cancer. 2016;97:1–7. doi: 10.1016/j.lungcan.2016.03.015. PubMed DOI

Vodicka J., Mukensnabl P., Vejvodova S., Spidlen V., Kulda V., Topolcan O., Pesta M. A more sensitive detection of micrometastases of NSCLC in lymph nodes using the one-step nucleic acid amplification (OSNA) method. J. Surg. Oncol. 2018;117:163–170. doi: 10.1002/jso.24826. PubMed DOI

Rusch V.W., Asamura H., Watanabe H., Giroux D.J., Rami-Porta R., Goldstraw P. The IASLC Lung Cancer Staging Project: A Proposal for a New International Lymph Node Map in the Forthcoming Seventh Edition of the TNM Classification for Lung Cancer. J. Thorac. Oncol. 2009;4:568–577. doi: 10.1097/JTO.0b013e3181a0d82e. PubMed DOI

Tamaki Y. One-step nucleic acid amplification (OSNA): Where do we go with it? Int. J. Clin. Oncol. 2017;22:3–10. doi: 10.1007/s10147-016-1030-9. PubMed DOI

Buglioni S., Di Filippo F., Terrenato I., Casini B., Gallo E., Marandino F., Maini C.L., Pasqualoni R., Botti C., Di Filippo S., et al. Quantitative molecular analysis of sentinel lymph node may be predictive of axillary node status in breast cancer classified by molecular subtypes. PLoS ONE. 2013;8:e58823. doi: 10.1371/journal.pone.0058823. PubMed DOI PMC

Tateishi M., Fukuyama Y., Hamatake M., Kohdono S., Ishida T., Sugimachi K. Skip mediastinal lymph node metastasis in non-small cell lung cancer. J. Surg. Oncol. 1994;57:139–142. doi: 10.1002/jso.2930570302. PubMed DOI

Wang L., Zhan C., Gu J., Xi J., Lin Z., Xue L., Ge D., Wang Q. Role of Skip Mediastinal Lymph Node Metastasis for Patients With Resectable Non-small-cell Lung Cancer: A Propensity Score Matching Analysis. Clin. Lung Cancer. 2019;20:e346–e355. doi: 10.1016/j.cllc.2018.12.007. PubMed DOI

Mehrpouya M., Pourhashem Z., Yardehnavi N., Oladnabi M. Evaluation of cytokeratin 19 as a prognostic tumoral and metastatic marker with focus on improved detection methods. J. Cell. Physiol. 2019;234:21425–21435. doi: 10.1002/jcp.28768. PubMed DOI

Noorlag R., van Es R.J.J., de Bree R., Willems S.M. Cytokeratin 19 expression in early oral squamous cell carcinoma and their metastasis: Inadequate biomarker for one-step nucleic acid amplification implementation in sentinel lymph node biopsy procedure: One-step nucleic acid amplification in early oral cancer. Head Neck. 2017;39:1864–1868. doi: 10.1002/hed.24847. PubMed DOI

Deambrogio C., Castellano I., Paganotti A., Zorini E.O., Corsi F., Bussone R., Franchini R., Antona J., Miglio U., Sapino A., et al. A new clinical cut-off of cytokeratin 19 mRNA copy number in sentinel lymph node better identifies patients eligible for axillary lymph node dissection in breast cancer. J. Clin. Pathol. 2014;67:702–706. doi: 10.1136/jclinpath-2014-202384. PubMed DOI

Terrenato I., D’Alicandro V., Casini B., Perracchio L., Rollo F., De Salvo L., Di Filippo S., Di Filippo F., Pescarmona E., Maugeri-Saccà M., et al. A cut-off of 2150 cytokeratin 19 mRNA copy number in sentinel lymph node may be a powerful predictor of non-sentinel lymph node status in breast cancer patients. PLoS ONE. 2017;12:e0171517. doi: 10.1371/journal.pone.0171517. PubMed DOI PMC

Ren Y., Zhang L., Xie H., She Y., Su H., Xie D., Zheng H., Zhang L., Jiang G., Wu C., et al. Lymph Node Micrometastasis Prognosticates Survival for Patients with Stage 1 Bronchogenic Adenocarcinoma. Ann. Surg. Oncol. 2018;25:3812–3819. doi: 10.1245/s10434-018-6743-8. PubMed DOI