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Background  Salivary rinses have been recently proposed as a valuable resource for the development of epigenetic biomarkers for detection and monitoring of head and neck squamous cell carcinoma (HNSCC). Both salivary rinses collected with and without an exfoliating brush from patients with HNSCC are used in detection of promoter hypermethylation, yet their correlation of promoter hypermethylation has not been evaluated. This study was to evaluate the concordance of promoter hypermethylation between salivary rinses collected with and without an exfoliating brush from patients with HNSCC.    Methodolgy  57 paired salivary rinses collected with or without an exfoliating brush from identical HNSCC patients were evaluated for promoter hypermethylation status using Quantitative Methylation-Specific PCR. Target tumor suppressor gene promoter regions were selected based on our previous studies describing a panel for HNSCC screening and surveillance, including  P16 ,  CCNA1 ,  DCC ,  TIMP3 ,  MGMT ,  DAPK  and  MINT31 .    Principal Findings  In salivary rinses collected with and without brush, frequent methylation was detected in  P16  (8.8% vs. 5.2%),  CCNA1  (26.3% vs. 22.8%),  DCC  (33.3% vs. 29.8%),  TIMP3  (31.6% vs. 36.8%),  MGMT  (29.8% vs. 38.6%),  DAPK  (14.0% vs. 19.2%), and  MINT31  (10.5% vs. 8.8%). Spearman's rank correlation coefficient showed a positive correlation between salivary rinses collected with and without brush for  P16  (ρ = 0.79),  CCNA1  (ρ = 0.61),  DCC  (ρ = 0.58),  TIMP3  (ρ = 0.10),  MGMT  (ρ = 0.70),  DAPK  (ρ = 0.51) and  MINT31  (ρ = 0.72) (P<0.01). The percent agreement of promoter methylation between salivary rinses with brush and without brush were 96.5% for  P16 , 82.5% for  CCNA1 , 78.9% for  DCC , 59.7% for  TIMP3 , 84.2% for  MGMT , 84.2% for  DAPK , and 94.7% for  MINT31 .    Conclusions  Our study demonstrated strong correlations of gene promoter hypermethylation between salivary rinses collected with and without an exfoliating brush. Salivary rinse collection without using an exfoliating brush may offer a cost effective, rapid, non-invasive, and reliable means for development of epigenetic salivary rinse biomarkers.

Comparison of Promoter Hypermethylation Pattern in Salivary Rinses Collected with and without an Exfoliating Brush from Patients with HNSCC