乳腺癌前哨淋巴结术中分子诊断的临床实用性初探

Evaluation of intraoperative molecular assay in sentinel lymph node biopsy from breast cancer patients

目的 探讨GeneSearchTM乳腺淋巴结检测试剂盒（以下简称GeneSearch）在乳腺癌前哨淋巴结（SLN）术中诊断的临床实用性.方法 对复旦大学附属肿瘤医院2009年2月至6月诊治的88例乳腺癌患者行SLN活检.首先垂直长轴将所得淋巴结切成数块厚约2 mm的组织块,对各切面进行术中细胞印片后,奇数号组织块用于术后连续切片检查,偶数号组织块采用GeneSearch进行检测,应用即时荧光定量逆转录聚合酶链反应检测SLN中CK19和乳腺球蛋白表达的Ct值.将GeneSearch以术后连续切片的诊断为准,与术中细胞印片、术后连续切片的病理结果分别进行比较.结果 88例共获得225枚SLN,其中宏转移淋巴结27枚,微转移淋巴结9枚,阴性淋巴结189枚（其中5枚为孤立肿瘤细胞）.从切割淋巴结开始到最终形成报告,GeneSearch耗时范围为35～45 min（平均40 min）.基于淋巴结数目,GeneSearch与术后连续切片的总体符合率为95.6%（215/225）,其检测敏感度为86.1%（31/36）,均高于术中细胞印片[分别为94.7%（213/225）和72.2%（26/36）].SLN转移灶大小与CKl9和乳腺球蛋白的Ct值存在统计学相关性（P＜0.01）.结论 GeneSearch用于SLN术中诊断时,其检测敏感度高于术中细胞印片,达到比较满意的效果,但在应用中仍存在一些问题.

Objective To evaluate the application of GeneSearchTM breast lymph node assay in intraoperative detection of metastases in sentinel lymph node （SLN） from breast cancer patients. Methods A total of 225 SLN from 88 patients was prospectively studied. Each SLN was cut into 2 mm slabs which were examined by intraoperative imprint cytology （ⅡC） first, followed by GeneSearch assay and postoperative serial sectioning. GeneSearch used real-time fluorescence quantitative RT-PCR technology to detect the expression of CK19 and mammaglobin in SLN. The results of GeneSearch assay were correlated with those of ⅡC and post-operative serial sectioning. Results Amongst the 88 cases studied, 225 SLNs were found, and obvious metastatic carcinoma cells were identified in 27 SLNs and micrometastasis in 9 SLNs.One hundred and eight-nine SLNs were considered as ＂negative＂ （with ＂isolated tumor cells＂ present in 5 SLNs）. The turn-around time of intraoperative GeneSearch assay ranged from 35 to 45 minutes （mean = 40 minutes）. The concordance rate between GeneSearch assay and post-operative serial sectioning was 95.6% （215/225）, with a sensitivity of 86.1% （31/36）, compared with 94.7% （213/225） and 72.2% （26/36）respectively for ⅡC. The size of metastatic foci correlated with the Ct value of CK19 and mammaglobin （P 〈0.01）. Conclusions GeneSearch assay for intraoperative detection of metastase in SLN has a satisfactory performance and demonstrates a relatively higher sensitivity than ⅡC. The potential clinical application still requires further evaluation of larger number of cases.