Abstract
Background
The purpose of this single-center study was to determine the practicability of the intra-operative use of one-step nucleic acid amplification (OSNA) as the only method for detection of SLN. The OSNA system has been well described and is supposed to be as accurate as conventional histology.
Methods
Three hundred and thirty SLNs from 143 breast cancer patients were analyzed in an intra-operative setting. The CK19-copy number was determined by OSNA and divided into 3 results (“−” no metastasis; “+” micrometastasis; “++” marcometastasis). If OSNA gave a positive result, an axillary lymph node dissection was carried out during the same session. The central 1-mm slice of each node was obtained for permanent histology. Additionally, the results were correlated to clinicopathological factors, and the time for the intra-operative use was evaluated.
Results
Thirty-nine of the 143 patients were OSNA positive, 22 with macrometastatic and 17 with micrometastatic spread. The mean time for the OSNA run with one SLN was 34.4 min. We could show a correlation between the tumor size and OSNA positivity as well as between the numbers of OSNA positive SLNs with the tumor load of associated non-SLNs. Furthermore, we found that a cutoff CK19 copy number of 7,900/μL indicates a positive non-SLN result with the highest sensitivity (91 %) and specificity (61 %).
Conclusion
We found OSNA to be very helpful for the intra-operative determination of the tumor load of a SLN as a basis for decision-making concerning further surgical axillary intervention. OSNA allows precise differentiation of micro- from macrometastasis, and the CK19 copy number predicts the probability of tumor load in other axillary lymph nodes and might help to find adequate adjuvant treatment options. This objective method is well suitable for everyday use and may reduce the pathologic workload and the risk of secondary operative interventions with all associated costs and stress for the patients.
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References
Bernet L, Cano R, Martinez M et al (2011) Diagnosis of the sentinel lymph node in breast cancer: a reproducible molecular method: a multicentric Spanish study. Histopathology 58(6):863–869
Chu PG, Weiss LM (2002) Keratin expression in human tissues and neoplasms. Histopathology 40(5):403–439. Review
Cserni G, Amendoeira I, Apostolikas N, Bellocq JP, Bianchi S, Boecker W, Borisch B, Connolly CE, Decker T, Dervan P, Drijkoningen M, Ellis IO, Elston CW, Eusebi V, Faverly D, Heikkila P, Holland R, Kerner H, Kulka J, Jacquemier J, Lacerda M, Martinez-Penuela J, De Miguel C, Peterse JL, Rank F, Regitnig P, Reiner A, Sapino A, Sigal-Zafrani B, Tanous AM, Thorstenson S, Zozaya E, Fejes G, Wells CA (2004) Discrepancies in current practice of pathological evaluation of sentinel lymph nodes in breast cancer. Results of a questionnaire based survey by the European Working Group for Breast Screening Pathology. J Clin Pathol 57(7):695–701
Cutress RI, McDowell A, Gabriel FG, Gill J, Jeffrey MJ, Agrawal A, Wise M, Raftery J, Cree IA, Yiangou C (2010) Observational and cost analysis of the implementation of breast cancer sentinel node intraoperative molecular diagnosis. J Clin Pathol 63(6):522–529
Espinosa-Bravo M, Sansano I, Pérez-Hoyos S, Ramos M, Sancho M, Xercavins J, Rubio IT, Peg V (2013) Prediction of non-sentinel lymph node metastasis in early breast cancer by assessing total tumoral load in the sentinel lymph node by molecular assay. Eur J Surg Onc 39(7):766–773 [Epub 2013 Apr 19]
Feldman S, Krishnamurthy S, Gillanders W, Gittleman M, Beitsch PD, Young PR, Streck CJ, Whitworth PW, Levine EA, Boolbol S, Han LK, Hermann R, Hoon DS, Giuliano AE, Meric-Bernstam F (2011) A novel automated assay for the rapid identification of metastatic breast carcinoma in sentinel lymph nodes. Cancer 117(12):2599–2607
Gillanders WE, Mikhitarian K, Hebert R et al (2004) Molecular detection of micrometastatic breast cancer in histopathology-negative axillary lymph nodes correlates with traditional predictors of prognosis: an interim analysis of a prospective multi-institutional cohort study. Ann Sur 239:828–837, discussion pp 837–840
Gimbergues P, Dauplat MM, Cayre A et al (2007) Correlation between molecular metastases in sentinel lymph nodes of breast cancer patients and St Gallen risk category. Eur J Surg Oncol 33:16–22
Goyal A, Newcombe RG, Chhabra A, Mansel RE (2008) Morbidity in breast cancer patients with sentinel node metastases undergoing delayed axillary lymph node dissection (ALND) compared with immediate ALND. Ann Surg Oncol 15(1):262–267
Guillén-Paredes MP, Carrasco-González L, Cháves-Benito A, Campillo-Soto A, Carrillo A, Aguayo-Albasini JL (2011) One-step nucleic acid amplification (OSNA) assay for sentinel lymph node metastases as an alternative to conventional postoperative histology in breast cancer: a cost-benefit analysis. Cir Esp 89(7):456–462
Klingler S, Marchal F, Rauch P, Kenouchi O, Chrétien AS, Genin P, Leroux A, Merlin JL (2013) Using one-step nucleic acid amplification (OSNA) for intraoperative detection of lymph node metastasis in breast cancer patients avoids second surgery and accelerates initiation of adjuvant therapy. Ann Oncol [Epub ahead of print]
Layfield DM, Agrawal A, Roche H, Cutress RI (2011) Intraoperative assessment of sentinel lymph nodes in breast cancer. Br J Surg 98(1):4–17
Le Frère-Belda MA, Bats AS, Gillaizeau F, Poulet B et al (2012) Diagnostic performance of one-step nucleic acid amplification for intraoperative sentinel node metastasis detection in breast cancer patients. Int J Cancer 130(10):2377–2386
Motumura K, Inaji H, Komoike Y et al (2000) Intraoperative lymph node examination by imprint cytology and frozen sectioning during breast surgery. Br J Surg 87:597–601
Ohi Y, Umekita Y, Sagara Y, Rai Y, Yotsumoto D, Matsukata A, Baba S, Tamada S, Matsuyama Y, Ando M, Sagara Y, Sasaki M, Tsuchimochi S, Tanimoto A, Sagara Y (2012) Whole sentinel lymph node analysis by a molecular assay predicts axillary node status in breast cancer. Br J Cancer 107(8):1239–1243
Olson JA Jr, McCall LM, Beitsch P, Whitworth PW, Reintgen DS, Blumencranz PW, Leitch AM, Saha S, Hunt KK, Giuliano AE, American College of Surgeons Oncology Group Trials Z0010 and Z0011 (2008) Impact of immediate versus delayed axillary node dissection on surgical outcomes in breast cancer patients with positive sentinel nodes: results from American College of Surgeons Oncology Group Trials Z0010 and Z0011. J Clin Oncol 26(21):3530–3535
Parikh RR, Yang Q, Higgins SA, Haffty BG (2008) Outcomes in young women with breast cancer of triple-negative phenotype: the prognostic significance of CK19 expression. Int J Radiat Oncol Biol Phys 70:35–42
Park D, Kåresen R, Naume B, Synnestvedt M, Beraki E, Sauer T (2009) The prognostic impact of occult nodal metastasis in early breast carcinoma. Breast Cancer Res Treat 118(1):57–66
Peg V, Espinosa-Bravo M, Vieites B, Vilardell F, Antúnez JR, de Salas MS, Delgado-Sánchez JJ, Pinto W, Gozalbo F, Petit A, Sansano I, Del Mar Téllez M, Rubio IT (2013) Intraoperative molecular analysis of total tumor load in sentinel lymph node: a new predictor of axillary status in early breast cancer patients. Brest Can Res Treat 139(1):87–93 [Epub 2013 Apr 11]
Roberts CA, Beitsch PD, Litz CE, Hilton DS, Ewing GE, Clifford E, Taylor W, Hapke MR, Babaian A, Khalid I, Hall JD, Lindberg G, Molberg K, Saboorian H (2003) Interpretive disparity among pathologists in breast sentinel lymph node evaluation. Am J Surg 186(4):324–329
Schem C, Maass N, Bauerschlag D et al (2009) One-step nucleic acid amplification—a molecular method of lymph node metastases in breast cancer patients; results of the German study group. Virchows Arch 454:203–210
Snook KL, Layer GT, Jackson PA, de Vries CS, Shousha S, Sinnett HD, Nigar E, Singhal H, Chia Y, Cunnick G, Kissin MW, OSNA Study Group (2011) Multicentre evaluation of intraoperative molecular analysis of sentinel lymph nodes in breast carcinoma. Br J Surg 98(4):527–535
Tamaki Y, Akiyama F, Iwase T et al (2009) Molecular detection of lymph node metastases in breast cancer patients: results of a multicenter trial using one-step nucleic acid amplification assay. Clin Can Res 15:2879–2884
Tew K, Irwig L, Matthews A, Crowe P, Macaskill P (2005) Meta-analysis of sentinel node imprint cytology in breast cancer. Br J Surg 92(9):1068–1080
Tsujimoto M, Nakabayashi K, Yoshidome K et al (2007) One-step nucleic acid amplification for intraoperative detection of lymph node metastasis in breast cancer patients. Clin Can Res 13:4807–4816
Van de Vrande S, Meijer J, Rijnders A, Klinkenbijl JH (2009) The value of intra operative frozen section examination of sentinel lymph nodes in breast cancer. Eur J SurgOncol 35(3):276–280
Vestjens JH, Pepels MJ, de Boer M, Borm GF, van Deurzen CH, van Diest PJ, van Dijck JA, Adang EM, Nortier JW, Rutgers EJ, Seynaeve C, Menke-Pluymers MB, Bult P, Tjan-Heijnen VC (2012) Relevant impact of central pathology review on nodal classification in individual breast cancer patients. Ann Oncol 23(10):2561–2566 [Epub 2012 Apr 11]
Vilardell F, Novell A, Martin J, Santacana M et al (2012) Importance of assessing CK19 immunostaining in core biopsies in patients subjected to sentinel node study by OSNA. Virchows Arch 460(6):569–575
Visser M, Jiwa M, Horstman A et al (2008) Intra-operative diagnostic method based on CK19 mRNA expression for the detection of lymph node metastases in breast cancer. Int J Cancer 122:2562–2567
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Thorsten Heilmann and Micaela Mathiak contributed equally to this project and should be considered co-first authors.
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Heilmann, T., Mathiak, M., Hofmann, J. et al. Intra-operative use of one-step nucleic acid amplification (OSNA) for detection of the tumor load of sentinel lymph nodes in breast cancer patients. J Cancer Res Clin Oncol 139, 1649–1655 (2013). https://doi.org/10.1007/s00432-013-1481-6
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DOI: https://doi.org/10.1007/s00432-013-1481-6
Keywords
- Carcinoma
- Breast cancer
- Sentinel lymph node/SLN
- Nucleic acid amplification/OSNA
- Tumor load
- CK19