One-step nucleic acid amplification for detecting lymph node metastasis of head and neck squamous cell carcinoma

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• 作者：Hiroyuki Goda ; Koh-ichi Nakashiro ; Ryota Oka ; Hiroshi Tanaka ; Hiroyuki Wakisaka ; Naohito Hato ; Masamitsu Hyodo ; Hiroyuki Hamakawa
• 关键词：One-step nucleic acid amplification (OSNA) ; Cytokeratin 19 (CK19) ; Head and neck squamous cell carcinoma (HNSCC) ; Lymph node metastasis ; Genetic diagnosis
• 刊名：Oral Oncology
• 出版年：2012
• 出版时间：October, 2012
• 年：2012
• 卷：48
• 期：10
• 页码：958-963
• 全文大小：349 K

文摘

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Summary

Objectives

Lymph node stage is an important prognostic factor in head and neck squamous cell carcinoma (HNSCC). We previously reported the clinical usefulness of sentinel lymph node biopsy diagnosed by genetic analysis using quantitative RT-PCR. However, this method takes about 3 h. In this study, we attempted to develop a more efficient method for the intraoperative genetic detection of lymph node metastasis in HNSCC.

Materials and methods

A total of 312 lymph nodes (65 patients) were diagnosed by the one-step nucleic acid amplification (OSNA) method using GD-100. OSNA consists of a short homogenization step followed by amplification of cytokeratin 19 (CK19) mRNA directly from the lysate. Each lymph node was divided into two to diagnose metastasis. One half was used for the OSNA assay, and the other was subjected to semi-serial sectioning, sliced at 200-¦Ìm intervals and examined by H&E and cytokeratin AE1/AE3 immunohistochemical staining. The accuracy of OSNA assay was evaluated based on histopathological diagnosis.

Results

Sixty-one of 312 lymph nodes were pathologically metastasis-positive. The overall concordance rate between the OSNA assay using breast cancer criteria and histopathology was 94.2 % . The optimal cut-off for the copy number of CK19 mRNA in assessing lymph node metastasis of HNSCC was 300 copies/¦Ìl, which had the highest diagnostic accuracy (95.2 % ). The OSNA assay can be completed within 30 min.

Conclusion

The OSNA assay, which shows high sensitivity and specificity, suggests the possibility to be used as a novel tool for the genetic detection of lymph node metastasis in HNSCC patients.