A pilot study to compare the detection of HPV-16 biomarkers in salivary oral rinses with tumour p16INK4a expression in head and neck squamous cell carcinoma patients

Abstract

BackgroundHuman papilloma virus-16 (HPV-16) infection is a major risk factor for a subset of head and neck squamous cell carcinoma (HNSCC), in particular oropharyngeal squamous cell carcinoma (OPSCC). Current techniques for assessing the HPV-16 status in HNSCC include the detection of HPV-16 DNA and p16INK4a expression in tumor tissues. When tumors originate from hidden anatomical sites, this method can be challenging. A non-invasive and cost-effective alternative to biopsy is therefore desirable for HPV-16 detection especially within a community setting to screen at-risk individuals.MethodsThe present study compared detection of HPV-16 DNA and RNA in salivary oral rinses with tumor p16INK4a status, in 82 HNSCC patients using end-point and quantitative polymerase chain reaction (PCR).ResultsOf 42 patients with p16INK4a-positive tumours, 39 (sensitivity = 92.9 %, PPV = 100 % and NPV = 93 %) had oral rinse samples with detectable HPV-16 DNA, using end-point and quantitative PCR. No HPV-16 DNA was detected in oral rinse samples from 40 patients with p16INK4a negative tumours, yielding a test specificity of 100 %. For patients with p16INK4a positive tumours, HPV-16 mRNA was detected using end-point reverse transcription PCR (RT-PCR) in 24/40 (sensitivity = 60 %, PPV = 100 % and NPV = 71 %), and using quantitative RT-PCR in 22/40 (sensitivity = 55 %, PPV = 100 % and NPV = 69 %). No HPV-16 mRNA was detected in oral rinse samples from the p16INK4a-negative patients, yielding a specificity of 100 %.ConclusionsWe demonstrate that the detection of HPV-16 DNA in salivary oral rinse is indicative of HPV status in HNSCC patients and can potentially be used as a diagnostic tool in addition to the current methods.Electronic supplementary materialThe online version of this article (doi:10.1186/s12885-016-2217-1) contains supplementary material, which is available to authorized users.

Oral testing for high‐risk human papillomavirus DNA and E6/E7 messenger RNA in healthy individuals at risk for oral infection

“…So far, the Aptima HPV assay has been used only to detect HPV mRNA in the saliva of patients with OPC . Another study reported HPV mRNA detection in oral rinses but used homemade, nonstandardized methods and included only patients with HNC …”

“…In contrast, HPV mRNA detection in oral specimens is very challenging because mRNA is highly susceptible to degradation by the enzymes present in oral fluids. Oral rinses, saliva, and swabs have been used only marginally for HPV mRNA detection in patients with OPC . To the best of our knowledge, no studies have reported HPV mRNA detection in oral rinses of cancer‐free individuals.…”

A Pilot Study into the Association between Oral Health Status and Human Papillomavirus—16 Infection

“…HPV-16-driven OPC constitute 90% of HPV-positive OPC [14]. The aetiology and risk factors of oral and oropharyngeal HPV infection remains controversial; however, past studies have shown that these risk factors include oral sex, multiple sexual partners, poor oral hygiene and immunodeficiency conditions [15,16,17,18,19,20]. Identifying key risk factors may help to improve disease screening processes to identify high-risk individuals for developing OPC which may lead to OPC prevention and improve disease mortality [21].…”

Rational genomic optimization of DNA detection for human papillomavirus type 16 in head and neck squamous cell carcinoma

“…Qureishi et al have reported successful detection of 72.2% HPV genome in their saliva samples obtained from HPV‐HNSCC cases. In a study by Chai et al, E6/E7 primer/probe sets showed 92.9% and 60% sensitivity for HPV‐16 genome in saliva samples for DNA and RNA, respectively. The data presented indicate that our combination of primer/probe sets would improve the sensitivity of detection of HPV16 DNA in tissues and body fluids and improve assessment of disease burden in patients with HPV‐HNSCC.…”