Combination Analysis of a Whole Lymph Node by One-Step Nucleic Acid Amplification and Histology for Intraoperative Detection of Micrometastasis

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<jats:p>Objective: The aim was to develop a more efficient molecular detection system than histological examination (HE) for lymph node (LN) metastasis. Methods: Cytokeratin (CK) 19 mRNA copy numbers of 5 colon carcinoma cell lines (Lovo, DLD1, WiDr, Colo201 and Colo320) were calculated and compared by one-step nucleic acid amplification (OSNA) and conventional real-time reverse-transcription polymerase chain reaction (RT-PCR). Then, 91 LN submitted for HE from 6 patients with advanced colorectal adenocarcinoma and 64 LN submitted for frozen diagnosis from 47 patients with different malignancies were examined by OSNA and HE. Results: CK19 mRNA copy numbers of all but Colo320 cells detected by OSNA were within double of those detected by RT-PCR. The least cell count of Lovo cells detected at one reaction (2 µl) by OSNA was calculated as 0.8 cells. Carcinoma metastasis showing either HE+ or OSNA+ was detected in 7.9% of the LN from advanced colorectal adenocarcinomas and in 30.0% of the LN for frozen diagnosis from different malignancies; HE–/OSNA+ metastasis was detected in 4.8 and 4.0%, respectively. OSNA analysis of 1 LN could be completed within 40 min. Conclusion: A combined analysis of LN by HE and OSNA could increase the sensitivity for detecting micrometastasis during surgery.</jats:p>

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  • Pathobiology

    Pathobiology 73 (4), 183-191, 2006

    S. Karger AG

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