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Poeta ML , Manola J , Goldenberg D , Forastiere A , Califano JA , Ridge JA , Goodwin J , Kenady D , Saunders J , Westra W , Sidransky D , Koch WM
The Ligamp TP53 Assay for Detection of Minimal Residual Disease in Head and Neck Squamous Cell Carcinoma Surgical Margins
Clinical Cancer Research. 2009 Dec;15(24) :7658-7665
PMID: ISI:000272853500027    PMCID: PMC2842212   
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Abstract
Purpose: Detect tumor-related DNA using LigAmp in histologically clear margins and associate results with clinical outcome. Experimental Design: 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 used two 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 and relative to tumor. Cutpoints were identified and defined groups were evaluated for local failure-free, cancer-specific, and overall survival. Study margins were examined for presence of tumor by light microscopy. Results: Tissue from 95 patients with common mutations was analyzed. Fifteen experienced local recurrence. Cutpoints of 0.15% for mutant species relative to plasmid and 0.5% for mutant species relative to tumor were chosen as most selective of recurrent cases. LigAmp had slightly better area under the receiver operator characteristic 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. Conclusions: 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. (Clin Cancer Res 2009;15(24):7658-65)
Notes
Poeta, M. Luana Manola, Judith Goldenberg, David Forastiere, Arlene Califano, Joseph A. Ridge, John A. Goodwin, Jarrard Kenady, Daniel Saunders, John Westra, William Sidransky, David Koch, Wayne M. NIH/NIDCR [R01 DE013152]; Public Health Service [CA23318, CA66636, CA21115, CA16116, CA27525]; National Cancer Institute ; Department of Health and Human Services NIH/NIDCR R01 DE013152. This study was coordinated by the Eastern Cooperative Oncology Group (Robert L. Comis, M.D., Chair) and supported in part by Public Health Service grants CA23318, CA66636, CA21115, CA16116, and CA27525 and from the National Cancer Institute, NIH and the Department of Health and Human Services. Its contents are solely the responsibility of the authors and do not necessarily represent the official views of the National Cancer Institute. 24 Amer assoc cancer research; 615 chestnut st, 17th floor, philadelphia, pa 19106-4404 usa 533vm