Annals of Surgical Oncology

, Volume 21, Issue 12, pp 3924–3930 | Cite as

The Diagnostic Value of One-Step Nucleic acid Amplification (OSNA) for Sentinel Lymph Nodes in Colon Cancer Patients

  • F. J. Vogelaar
  • M. S. Reimers
  • R. L. A. van der Linden
  • J. C. van der Linden
  • V. T. H. B. M. Smit
  • D. J. Lips
  • C. J. H. van de Velde
  • K. BosschaEmail author
Colorectal Cancer



Lymph node status in colon cancer is critical for prognosis estimation and treatment allocation. The purpose of this study was to compare the performance of one-step nucleic acid amplification (OSNA) through detection of cytokeratin 19 mRNA levels with routine pathological examination (RP) and multilevel fine pathological examination (FP) in sentinel lymph nodes (SLN), detected using the ex vivo SLN mapping (SLNM) procedure, in presurgically defined nonmetastatic colon cancer patients.


In this prospective study, 325 SLNs of 128 patients from the Jeroen Bosch Hospital in ‘s-Hertogenbosch and the Leiden University Medical Center were investigated by RP (H&E), FP (H&E and Keratin Pan immunohistochemical staining), and OSNA. The SLNs were harvested by the SLNM procedure, using Patent blue or Indocyanine green. SLNs were divided and separate parts were used for RP, FP, and the OSNA assay.


The diagnostic value of OSNA was 82.1 and 100 % for both FP and combined method (OSNA and FP) compared with RP. An upstaging rate of 20.2 % was obtained with the use of OSNA only and 36.4 % with the use of FP only. An upstaging rate of 46.5 % was obtained by combining the two methods together.


OSNA and FP appeared to be promising tools for the detection of lymph node micro- and macrometastases in SLNs after SLNM. The performances of OSNA and FP in this study were superior to RP. Because OSNA allows analysis of the whole lymph node, sampling bias can be avoided. OSNA therefore may improve tumor staging.


Sentinel Lymph Node Fine Pathological Isolate Tumor Cell Sentinel Lymph Node Mapping Routine Pathological 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.



The authors thank the image-guided surgery group from the LUMC for their help and expertise in the detection of SLNs, Dr. Liefers for advise and data interpretation, Dr. Takata for the study design and data interpretation, M. Hilbink for statistical advise, J. Damen and N.ter Haar for technical support, and Sysmex Corporation for technical support and providing study material.



Supplementary material

10434_2014_3820_MOESM1_ESM.pdf (84 kb)
Supplementary material 1 (PDF 84 kb)


  1. 1.
  2. 2.
    Hermanek P. Prognostic factor research in oncology. J Clin Epidemiol. 1999;52:371–4.PubMedCrossRefGoogle Scholar
  3. 3.
    Radespiel-Troger M, Hohenberger W, Reingruber B. Improved prediction of recurrence after curative resection of colon carcinoma using tree-based risk stratification. Cancer. 2004;100:958–67.PubMedCrossRefGoogle Scholar
  4. 4.
    Tepper JE, Wang AZ. Improving local control in rectal cancer: radiation sensitizers or radiation dose? J Clin Oncol. 2010;28:1623–4.PubMedCrossRefGoogle Scholar
  5. 5.
    Cianchi F, Palomba A, Boddi V, et al. Lymph node recovery from colorectal tumor specimens: recommendation for a minimum number of lymph nodes to be examined. World J Surg. 2002;26:384–9.PubMedCrossRefGoogle Scholar
  6. 6.
    Leibl S, Tsybrovskyy O, Denk H. How many lymph nodes are necessary to stage early and advanced adenocarcinoma of the sigmoid colon and upper rectum? Virchows Arch. 2003;443:133–8.PubMedCrossRefGoogle Scholar
  7. 7.
    Yoshimatsu K, Ishibashi K, Umehara A, et al. How many lymph nodes should be examined in Dukes’ B colorectal cancer? Determination on the basis of cumulative survival rate. Hepatogastroenterology. 2005;52:1703–6.PubMedGoogle Scholar
  8. 8.
    Koebrugge B, Vogelaar FJ, Lips DJ, et al. The number of high-risk factors is related to outcome in stage II colonic cancer patients. Eur J Surg Oncol. 2011;37:964–70.PubMedCrossRefGoogle Scholar
  9. 9.
    Burke HB. Outcome prediction and the future of the TNM staging system. J Natl Cancer Inst. 2004;96:1408–9.PubMedCrossRefGoogle Scholar
  10. 10.
    Feezor RJ, Copeland EM III, Hochwald SN. Significance of micrometastases in colorectal cancer. Ann Surg Oncol. 2002;9:944–53.PubMedCrossRefGoogle Scholar
  11. 11.
    O’Connell JB, Maggard MA, Ko CY. Colon cancer survival rates with the new American Joint Committee on Cancer sixth edition staging. J Natl Cancer Inst. 2004;96:1420–5.PubMedCrossRefGoogle Scholar
  12. 12.
    Yamamoto H, Sekimoto M, Oya M, et al. OSNA-based novel molecular testing for lymph node metastases in colorectal cancer patients: results from a multicenter clinical performance study in Japan. Ann Surg Oncol. 2011;18:1891–8.PubMedCrossRefGoogle Scholar
  13. 13.
    Liefers GJ, Cleton-Jansen AM, van de Velde CJ, et al. Micrometastases and survival in stage II colorectal cancer. N Engl J Med. 1998;339:223–8.PubMedCrossRefGoogle Scholar
  14. 14.
    van Schaik PM, Hermans E, van der Linden JC, et al. Micro-metastases in stages I and II colon cancer are a predictor of the development of distant metastases and worse disease-free survival. Eur J Surg Oncol. 2009;35:492–6.PubMedCrossRefGoogle Scholar
  15. 15.
    Viehl CT, Guller U, Cecini R, et al. Sentinel lymph node procedure leads to upstaging of patients with resectable colon cancer: results of the Swiss prospective, multicenter study sentinel lymph node procedure in colon cancer. Ann Surg Oncol. 2012;19:1959–65.PubMedCrossRefGoogle Scholar
  16. 16.
    Tsujimoto M, Nakabayashi K, Yoshidome K, et al. One-step nucleic acid amplification for intraoperative detection of lymph node metastasis in breast cancer patients. Clin Cancer Res. 2007;13:4807–16.PubMedCrossRefGoogle Scholar
  17. 17.
    Visser M, Jiwa M, Horstman A, et al. Intraoperative rapid diagnostic method based on CK19 mRNA expression for the detection of lymph node metastases in breast cancer. Int J Cancer. 2008;122:2562–7.PubMedCrossRefPubMedCentralGoogle Scholar
  18. 18.
    Tamaki Y, Akiyama F, Iwase T, et al. Molecular detection of lymph node metastases in breast cancer patients: results of a multicenter trial using the one-step nucleic acid amplification assay. Clin Cancer Res. 2009;15:2879–84.PubMedCrossRefGoogle Scholar
  19. 19.
    Croner RS, Schellerer V, Demund H, et al. One step nucleic acid amplification (OSNA)—a new method for lymph node staging in colorectal carcinomas. J Transl Med. 2010;8:83.PubMedCrossRefPubMedCentralGoogle Scholar
  20. 20.
    Guller U, Zettl A, Worni M, et al. Molecular investigation of lymph nodes in colon cancer patients using one-step nucleic acid amplification (OSNA): a new road to better staging? Cancer. 2012;118:6039–45.PubMedCrossRefPubMedCentralGoogle Scholar
  21. 21.
    Bilchik AJ, Nora D, Tollenaar RA, et al. Ultrastaging of early colon cancer using lymphatic mapping and molecular analysis. Eur J Cancer. 2002;38:977–85.PubMedCrossRefGoogle Scholar
  22. 22.
    Cahill RA, Leroy J, Marescaux J. Could lymphatic mapping and sentinel node biopsy provide oncological providence for local resectional techniques for colon cancer? A review of the literature. BMC Surg. 2008;8:17.PubMedCrossRefPubMedCentralGoogle Scholar
  23. 23.
    Lips DJ, Koebrugge B, Liefers GJ, et al. The influence of micrometastases on prognosis and survival in stage I-II colon cancer patients: the Enroute plus sign in circle Study. BMC Surg. 2011;11:11.PubMedCrossRefPubMedCentralGoogle Scholar
  24. 24.
    Schaafsma BE, Verbeek FP, van der Vorst JR, et al. Ex vivo sentinel node mapping in colon cancer combining blue dye staining and fluorescence imaging. J Surg Res. 2013;183:253–7.PubMedCrossRefGoogle Scholar
  25. 25.
    Taniyama K, Motoshita J, Sakane J, et al. Combination analysis of a whole lymph node by one-step nucleic acid amplification and histology for intraoperative detection of micrometastasis. Pathobiology. 2006;73:183–91.PubMedCrossRefGoogle Scholar
  26. 26.
    Bilchik A, Nissan A, Wainberg Z, et al. Surgical quality and nodal ultrastaging is associated with long-term disease-free survival in early colorectal cancer: an analysis of 2 international multicenter prospective trials. Ann Surg. 2010;252:467–74.PubMedGoogle Scholar
  27. 27.
    Rosenberg R, Friederichs J, Gertler R, et al. Prognostic evaluation and review of immunohistochemically detected disseminated tumor cells in peritumoral lymph nodes of patients with pN0 colorectal cancer. Int J Colorectal Dis. 2004;19:430–7.PubMedCrossRefGoogle Scholar
  28. 28.
    Richtlijnen oncologische zorg. 2014. Online source.
  29. 29.
    Yamamoto H, Sekimoto M, Oya M, et al. OSNA-based novel molecular testing for lymph node metastases in colorectal cancer patients: results from a multicenter clinical performance study in Japan. Ann Surg Oncol. 2011;18:1891–8.PubMedCrossRefGoogle Scholar
  30. 30.
    Bilchik AJ, Hoon DS, Saha S, et al. Prognostic impact of micrometastases in colon cancer: interim results of a prospective multicenter trial. Ann Surg. 2007;246:568–75.PubMedCrossRefGoogle Scholar
  31. 31.
    Bosch Roig CE, Rosello-Sastre E, Alonso HS, et al. Prognostic value of the detection of lymph node micrometastases in colon cancer. Clin Transl Oncol. 2008;10:572–8.Google Scholar
  32. 32.
    Faerden AE, Sjo OH, Bukholm IR, et al. Lymph node micrometastases and isolated tumor cells influence survival in stage I and II colon cancer. Dis Colon Rectum. 2011;54:200–6.PubMedCrossRefGoogle Scholar
  33. 33.
    Park SJ, Lee KY, Kim SY. Clinical significance of lymph node micrometastasis in stage I and II colon cancer. Cancer Res Treat. 2008;40:75–80.PubMedCrossRefPubMedCentralGoogle Scholar

Copyright information

© Society of Surgical Oncology 2014

Authors and Affiliations

  • F. J. Vogelaar
    • 1
    • 3
  • M. S. Reimers
    • 4
  • R. L. A. van der Linden
    • 2
  • J. C. van der Linden
    • 2
  • V. T. H. B. M. Smit
    • 5
  • D. J. Lips
    • 1
  • C. J. H. van de Velde
    • 4
  • K. Bosscha
    • 1
    Email author
  1. 1.Department of SurgeryJeroen Bosch Hospital‘s-HertogenboschThe Netherlands
  2. 2.Department of PathologyJeroen Bosch Hospital‘s-HertogenboschThe Netherlands
  3. 3.Department of SurgeryVieCuri Medical CenterVenloThe Netherlands
  4. 4.Department of SurgeryLeiden University Medical CenterLeidenThe Netherlands
  5. 5.Department of PathologyLeiden University Medical CenterLeidenThe Netherlands

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