J Breast Cancer.  2010 Dec;13(4):366-374. 10.4048/jbc.2010.13.4.366.

One-step Nucleic Acid Amplification (OSNA): Intraoperative Rapid Molecular Diagnostic Method for the Detection of Sentinel Lymph Node Metastases in Breast Cancer Patients in Korean Cohort

• Choi YL
• Ahn SK
• Bae YK
• Park IA
• Min JW
• Lee KW
• Bae JH
• Han W
• Choi JE
• Cho EY
• Kim S
• Lee HS
• Nam SJ
• Lee JE
• Lee SJ
• Noh DY
• Yang JH

• KMID: 2286524
• DOI: http://doi.org/10.4048/jbc.2010.13.4.366

Abstract

PURPOSE
Sentinel lymph node (SLN) biopsy has become a standard procedure in breast cancer patient management. Accurate intraoperative assessment of metastasis of SLNs is essential for appropriate selection to avoid unnecessary axillary dissection. The aim of this study was to evaluate the performance of one-step nucleic acid amplification (OSNA) assay for detection of sentinel lymph node metastasis examination in breast cancer patients.
METHODS
In this study, we compared intraoperative OSNA to histological investigation with multi-level observation in 284 sentinel lymph nodes of 199 patients. Surgically obtained sentinel lymph nodes were sectioned into 2 mm intervals of up to four pieces, half of which were examined with the OSNA assay. The other half of adjacent pieces were histopathologically examined both intraoperatively and postoperatively. The presence/absence of metastases was judged by observing hematoxylin and eosin staining and cytokeratin (AE1/ AE3) immunohistochemically stained multiple slides from one lymph node.
RESULTS
Among 199 patients included, 36 cases were positive on histological examination and 34 cases were positive on OSNA assay. There were 14 discordant cases. The overall concordance with histology was 93.0% (95% confidence interval [CI], 0.86-0.96), with a sensitivity of 77.8% (95% CI, 0.61-0.90), specificity of 96.3% (95% CI, 0.92-0.99), positive predictive value of 82.4% (95% CI, 0.65-0.93) and negative predictive value of 95.2% (95% CI, 0.91-0.98). The kappa statistic analysis indicated substantial agreement of both methods, with a value of 0.76 (95% CI, 0.64-0.88). The average turnaround time was 39.0 minutes.
CONCLUSION
The results of this study indicate that the OSNA assay has equivalent accuracy to histopathology in detecting breast cancer metastasis to lymph nodes when each method is assigned two alternate blocks of four blocks sectioned at 2 mm intervals.

Keyword

Breast neoplasms; Molecular diagnostic techniques; Sentinel lymph node biopsy

MeSH Terms

Biopsy
Breast
Breast Neoplasms
Cohort Studies
Eosine Yellowish-(YS)
Hematoxylin
Humans
Keratins
Lymph Nodes
Molecular Diagnostic Techniques
Neoplasm Metastasis
Nitriles
Pathology, Molecular
Pyrethrins
Sensitivity and Specificity
Sentinel Lymph Node Biopsy
Eosine Yellowish-(YS)
Hematoxylin
Keratins
Nitriles
Pyrethrins

Figure

• 

  Figure 1 (A) Resected lymph nodes sectioned at 2 mm intervals. Blocks a and c were subjected to one-step nucleic acid amplification (OSNA) assay, and blocks b and d to histopathological examination. (B) Frozen sections for H&E staining prepared from one level of two cutting surfaces of blocks b and d. (1 [red] and 3 [red]). After being formalin-fixed and paraffin-embedded, one cut-surface of block b and one cut-surface of block d, and three level sections (green) at 200 µm interval of specimens for histopathological examination were prepared. Three specimens were prepared for each level: H&E staining, anti-cytokeratin antibody staining and unstained.

Reference

1. Korean Breast Cancer Society. Breast Cancer Facts and Figures 2006-2008. Vol. 1. 2008. Seoul: Korean Breast Cancer Society;5–7.

2. Giuliano AE, Kirgan DM, Guenther JM, Morton DL. Lymphatic mapping and sentinel lymphadenectomy for breast cancer. Ann Surg. 1994. 220:391–398.

3. Giuliano AE, Dale PS, Turner RR, Morton DL, Evans SW, Krasne DL. Improved axillary staging of breast cancer with sentinel lymphadenectomy. Ann Surg. 1995. 222:394–399.

4. Mabry H, Giuliano AE. Sentinel node mapping for breast cancer: progress to date and prospects for the future. Surg Oncol Clin N Am. 2007. 16:55–70.

5. Sung WJ, Kim A, Kang SH, Lee SJ, Hwang TY, Bae YK. The usefulness and limitations of intraoperative frozen section analysis of sentinel lymph nodes in patients with breast cancer. J Breast Cancer. 2009. 12:170–178.

6. Veronesi U, Paganelli G, Viale G, Galimberti V, Luini A, Zurrida S, et al. Sentinel lymph node biopsy and axillary dissection in breast cancer: results in a large series. J Natl Cancer Inst. 1999. 91:368–373.

7. D'Errico A, Grassigli A, Gruppioni E, Fiorentino M, Corti B, Gabusi E, et al. Thorough intraoperative analysis of breast sentinel lymph node biopsies: histologic and immunohistochemical findings. Surgery. 2004. 135:248–254.

8. Edge SB, Byrd DR, Compton CC, Fritz AG, Greene FL, Trotti A. AJCC Cancer Staging Manual. 2010. New York: Springer-Verlag;347–377.

9. Tsujimoto M, Nakabayashi K, Yoshidome K, Kaneko T, Iwase T, Akiyama F, et al. One-step nucleic acid amplification for intraoperative detection of lymph node metastasis in breast cancer patients. Clin Cancer Res. 2007. 13:4807–4816.

10. Visser M, Jiwa M, Horstman A, Brink AA, Pol RP, van Diest P, et al. Intra-operative rapid diagnostic method based on CK19 mRNA expression for the detection of lymph node metastases in breast cancer. Int J Cancer. 2008. 122:2562–2567.

11. Schem C, Maass N, Bauerschlag DO, Carstensen MH, Löning T, Roder C, et al. One-step nucleic acid amplification-a molecular method for the detection of lymph node metastases in breast cancer patients: results of the German study group. Virchows Arch. 2009. 454:203–210.

12. Tamaki Y, Akiyama F, Iwase T, Kaneko T, Tsuda H, Sato K, 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. 15:2879–2884.

13. Nagamine K, Hase T, Notomi T. Accelerated reaction by loop-mediated isothermal amplification using loop primers. Mol Cell Probes. 2002. 16:223–229.

14. Singletary SE, Greene FL. Breast Task Force. Revision of breast cancer staging: the 6th edition of the TNM Classification. Semin Surg Oncol. 2003. 21:53–59.

15. Langer I, Guller U, Berclaz G, Koechli OR, Moch H, Schaer G, et al. Accuracy of frozen section of sentinel lymph nodes: a prospective analysis of 659 breast cancer patients of the Swiss multicenter study. Breast Cancer Res Treat. 2009. 113:129–136.

16. van de Vrande S, Meijer J, Rijnders A, Klinkenbijl JH. The value of intraoperative frozen section examination of sentinel lymph nodes in breast cancer. Eur J Surg Oncol. 2009. 35:276–280.

17. Schwartz GF, Krill LS, Palazzo JP, Dasgupta A. Value of intraoperative examination of axillary sentinel nodes in carcinoma of the breast. J Am Coll Surg. 2008. 207:758–762.

18. Ali R, Hanly AM, Naughton P, Castineira CF, Landers R, Cahill RA, et al. Intraoperative frozen section assessment of sentinel lymph nodes in the operative management of women with symptomatic breast cancer. World J Surg Oncol. 2008. 6:69–74.

19. Choi YJ, Yun HR, Yoo KE, Kim JH, Nam SJ, Choi YL, et al. Intraoperative examination of sentinel lymph nodes by ultrarapid immunohistochemistry in breast cancer. Jpn J Clin Oncol. 2006. 36:489–493.

20. Weiser MR, Montgomery LL, Susnik B, Tan LK, Borgen PI, Cody HS. Is routine intraoperative frozen-section examination of sentinel lymph nodes in breast cancer worthwhile? Ann Surg Oncol. 2000. 7:651–655.

21. Backus J, Laughlin T, Wang Y, Belly R, White R, Baden J, et al. Identification and characterization of optimal gene expression markers for detection of breast cancer metastasis. J Mol Diagn. 2005. 7:327–336.

22. Mitas M, Mikhitarian K, Walters C, Baron PL, Elliott BM, Brothers TE, et al. Quantitative real-time RT-PCR detection of breast cancer micrometastasis using a multigene marker panel. Int J Cancer. 2001. 93:162–171.

23. Noguchi S, Aihara T, Nakamori S, Motomura K, Inaji H, Imaoka S, et al. The detection of breast carcinoma micrometastases in axillary lymph nodes by means of reverse transcriptase-polymerase chain reaction. Cancer. 1994. 74:1595–1600.

24. Mansel RE, Goyal A, Douglas-Jones A, Woods V, Goyal S, Monypenny I, et al. Detection of breast cancer metastasis in sentinel lymph nodes using intra-operative real time GeneSearch BLN Assay in the operating room: results of the Cardiff study. Breast Cancer Res Treat. 2009. 115:595–600.

25. Bartek J, Taylor-Papadimitriou J, Miller N, Millis R. Patterns of expression of keratin 19 as detected with monoclonal antibodies in human breast tissues and tumours. Int J Cancer. 1985. 36:299–306.