Combination analysis of a whole lymph node by one-step nucleic acid amplification and histology for intraoperative detection of micrometastasis
Combination analysis of a whole lymph node by one-step nucleic acid amplification and histology for intraoperative detection of micrometastasis
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DOI：
10.1159/000096019
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发表时间：
2006-01-01
期刊：
影响因子：
5
通讯作者：
Koseki, Masato
中科院分区：
文献类型：
--
作者：
Taniyama, Kiyomi;Motoshita, Junichi;Koseki, Masato
目的：建立一种比组织学检查（HE）更有效的淋巴结（LN）转移分子检测系统。研究方法：应用一步核酸扩增法（C SNA）和实时荧光定量逆转录聚合酶链反应（RT-PCR）检测5种结肠癌细胞系（Lovo、DLD 1、WiDr、Colo 201和Colo 320）细胞角蛋白（CK）19 mRNA拷贝数。对6例晚期结直肠腺癌患者的91例LN和47例不同恶性肿瘤患者的64例LN进行组织学检查。结果：除Colo 320细胞外，其他细胞CK 19 mRNA拷贝数均在RT-PCR的2倍以内。通过OSNA在一个反应（2 μ l）中检测到的Lovo细胞的最少细胞计数计算为0.8个细胞。在晚期结直肠腺癌的LN中检测到7.9%的显示HE+或OSNA+的癌转移，在不同恶性肿瘤的冷冻诊断的LN中检测到30.0%的癌转移;分别在4.8%和4.0%中检测到HE-/OSNA+转移。1例LN的OSNA分析可在40 min内完成。结论：HE和OSNA联合分析LN可提高术中微转移检测的敏感性。版权所有（c）2006 S. Karger AG，巴塞尔。
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 (C)SNA) 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 mu l) 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. Copyright (c) 2006 S. Karger AG, Basel.