Abstract
Purpose
HPV-related oropharyngeal squamous cell carcinomas clearly represent a growing entity in the head and neck with distinct carcinogenesis, clinico-pathological presentation and survival profile. We aimed to compare the HPV prevalence rates and clinico-pathological correlations obtained with three distinct commonly used HPV detection methods.
Materials and Methods
p16-immunohistochemistry (IHC), HPV DNA viral load by real-time PCR (qPCR), and HPV genotyping by a reverse hybridization-based line probe assay (INNO-LiPA) were performed on pretreatment formalin-fixed paraffin-embedded tumor samples from 46 patients treated for single primary oropharyngeal carcinomas.
Results
Twenty-eight patients (61%) had a p16 overexpression in IHC. Twenty-nine patients (63%) harbored HPV DNA on qPCR. Thirty-four patients (74%) harbored HPV DNA on INNO-LiPA. The concordance analysis revealed a good agreement between both HPV DNA detection methods (κ = 0.65); when both tests were positive, the depicted HPV subtypes were always concordant (HPV16 in 27 cases, HPV18 in 1 case). Agreement was moderate between IHC and qPCR (κ = 0.59) and fair between IHC and INNO-LiPA (κ = 0.22).
Conclusions
Certain highly sensitive methods are able to detect the mere presence of HPV without any carcinogenetic involvement while other more specific tests provide proof of viral transcriptional activity and thus evidence of clinically relevant infections. The use of a stepwise approach allows reducing false positives; p16-immunostaining seems to be an excellent screening test and in situ hybridization may overcome some of the PCR limitations.
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Introduction
Head and neck squamous cell carcinomas (HNSCC) are the sixth most common cancers worldwide, accounting for 633000 new cases annually . In western countries, a significant regression of their incidence has been observed over the past 30 years, mostly explained by a decrease in tobacco and ethanol exposures. This overall decrease was paradoxically associated to an increase in incidence of oropharyngeal squamous cell carcinomas (OPSCC), especially in young patients with no known carcinogen exposure, in whom the role of the high-risk human papillomavirus (HR-HPV) infection is now clearly established . Numerous reports are in agreement with the fact that the HPV-related subset of OPSCC represents a clinically distinct disease with a different carcinogenesis and a more favorable survival profile than its stage-matched non-HPV-related counterpart, and may therefore benefit from different therapeutic approaches .
Although systematic HPV testing has been widely recommended in OPSCC, there is no consensual detection method yet . A wide spectrum of detection techniques is currently available ; the information they provide is however largely variable, which makes the reliability of results questionable, and their comparison among different series somehow restricted. In the present study, we aimed to compare the HPV prevalence rates and clinico-pathological correlations obtained with three distinct commonly used HPV detection methods: p16 INK4A expression status by immunohistochemistry (IHC), HPV DNA viral load by quantitative real-time PCR (qPCR), and HPV genotyping by a reverse hybridization-based line probe assay (LiPA; the INNO-LiPA). We also reviewed the currently available HPV detection techniques and outlined their major advantages and drawbacks.
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