The Ligamp TP53 Assay for Detection of Minimal Residual Disease in Head and Neck Squamous Cell Carcinoma Surgical Margins.
The Ligamp TP53 Assay for Detection of Minimal Residual Disease in Head and Neck Squamous Cell Carcinoma Surgical Margins.
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DOI：
10.1158/1078-0432.ccr-09-1433
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发表时间：
2009-12-15
期刊：
影响因子：
--
通讯作者：
Koch WM
中科院分区：
文献类型：
--
作者：
Poeta ML;Manola J;Goldenberg D;Forastiere A;Califano JA;Ridge JA;Goodwin J;Kenady D;Saunders J;Westra W;Sidransky D;Koch WM
在组织学清晰的边缘使用LigAmp检测肿瘤相关DNA，并将结果与临床结果相关联。头颈癌患者登记进行手术边缘分子分析。利用组织学边缘分析确保切除的充分性。然后收集更多的边缘并提取DNA。采用Affymetrix p53基因芯片检测肿瘤中TP53突变。用Ligamp对边缘进行分析，并与定量突变DNA的标准曲线进行比较。结扎采用2个寡核苷酸分离靶向突变的DNA。用实时PCR扩增结扎DNA。边缘突变的数量以突变种相对于质粒（MRP）和相对于肿瘤（MRT）的百分比来确定。确定切点并定义组，评估局部无失败、癌症特异性和总生存期。光镜检查研究边缘是否存在肿瘤。分析了95例常见突变患者的组织。15例局部复发。MRP的截点为0.15%，MRT的截点为0.5%，这是最具选择性的复发病例。LigAmp在ROC曲线下的面积（p=0.09）略优于光学显微镜，正确预测了15例复发肿瘤中的9例。假阴性6例，假阳性26例。在这个有限的队列中，在癌症特异性或总生存率方面没有观察到统计学上显著的差异。Ligamp提供了可量化的，敏感的检测突变DNA在组织学上正常的边缘。检测边缘的突变物种可以识别有局部复发风险的患者。
Detect tumor-related DNA using LigAmp in histologically clear margins and associate results with clinical outcome. Patients with head and neck cancer were registered for molecular analysis of surgical margins. Adequacy of resection was ensured using histologic margin analysis. Further margins were then harvested and DNA extracted. TP53 mutations in tumor were determined using Affymetrix p53 GeneChip. Margins were analyzed by Ligamp in comparison with standard curves for quantification of mutant DNA. Ligation employed 2 oligonucleotides to isolate DNA targeting the mutation. Ligated DNA was amplified using real-time PCR. The quantity of mutation in the margin was determined as percent of mutant species relative to plasmid (MRP) and relative to tumor (MRT). Cutpoints were identified and defined groups evaluated for local failure-free, cancer-specific, and overall survival. Study margins were examined for presence of tumor by light microscopy. Tissue from 95 patients with common mutations was analyzed. Fifteen experienced local recurrence. Cutpoints of 0.15% for MRP and 0.5% for MRT were chosen as most selective of recurrent cases. LigAmp had slightly better area under the ROC curve (p=0.09) than light microscopy correctly predicting 9 of 15 recurrent tumors. There were 6 false negative cases and 26 false positive results. No statistically significant distinctions were observed in cancer-specific or overall survival in this limited cohort. Ligamp provides quantifiable, sensitive detection of mutant DNA in histologically normal margins. Detection of mutant species in margins may identify patients at risk of local recurrence.