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Publication
p16 Immunohistochemistry in Oropharyngeal Squamous Cell Carcinoma Using the E6H4
Antibody Clone: A Technical Method Study for Optimal Dilution.
Authors Lewis JS, Shelton J, Kuhs KL, K Smith D
Submitted By Submitted Externally on 12/12/2018
Status Published
Journal Head and neck pathology
Year 2018
Date Published 12/1/2018
Volume : Pages 12 : 440 - 447
PubMed Reference 29190003
Abstract Routine testing for p16 immunohistochemistry (with selective HPV-specific test
use) has been recommended for clinical practice in oropharyngeal squamous cell
carcinoma (OPSCC). Data suggests that the E6H4 clone performs best for this
purpose, yet no studies have evaluated the optimal antibody concentration for
OPSCC testing. We evaluated three concentrations (undiluted, 1:5, and 1:10) of
the primary antibody solution for E6H4 using tissue microarrays from a cohort of
199 OPSCC patients with a?>?70% staining cutoff for positivity. Concordance was
evaluated using percent agreement and Cohen's kappa. The concentrations were
evaluated for sensitivity and specificity using high risk HPV RNA in situ
hybridization (RNA-ISH) and also correlated with Kaplan-Meier overall survival
analysis. Inter-rater agreement was very high between p16 results at each
concentration and also with RNA in situ hybridization (p?Agreement between p16 undiluted and 1:5 dilution (agreement 98.2%; Kappa 0.943;
p?79.2%; Kappa 0.512; p?decrease with the 1:5 and 1:10 dilutions compared to undiluted, but not in a
manner that obviously would change test interpretation or performance. Results
suggest that the E6H4 antibody performs well at dilutions of up to 1:5 fold with
a minor decrease in staining intensity, minimum loss of sensitivity, and no loss
of specificity in OPSCC patients. This could result in reagent and cost savings.




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