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. 2016 Mar 3:16:178.
doi: 10.1186/s12885-016-2217-1.

A pilot study to compare the detection of HPV-16 biomarkers in salivary oral rinses with tumour p16(INK4a) expression in head and neck squamous cell carcinoma patients

Ryan C Chai  1   2 , Yenkai Lim  3 , Ian H Frazer  4 , Yunxia Wan  5 , Christopher Perry  6 , Lee Jones  7 , Duncan Lambie  8 , Chamindie Punyadeera  9
Affiliations

A pilot study to compare the detection of HPV-16 biomarkers in salivary oral rinses with tumour p16(INK4a) expression in head and neck squamous cell carcinoma patients

Ryan C Chai et al. BMC Cancer. .

Abstract

Background: Human 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 p16(INK4a) 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.

Methods: The present study compared detection of HPV-16 DNA and RNA in salivary oral rinses with tumor p16(INK4a) status, in 82 HNSCC patients using end-point and quantitative polymerase chain reaction (PCR).

Results: Of 42 patients with p16(INK4a)-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 p16(INK4a) negative tumours, yielding a test specificity of 100 %. For patients with p16(INK4a) 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 p16(INK4a)-negative patients, yielding a specificity of 100 %.

Conclusions: We 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.

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Figures

Fig. 1
Fig. 1
HPV-16 DNA in oral rinse associates with tumor p16INK4a status. a Detection of HPV-16 DNA in representative oral rinse samples of patients with p16INK4a-positive and p16INK4a-negative tumors using end-point PCR. (b) Analysis of HPV-16 DNA copy number in oral rinse of patients with p16INK4a-positive tumors using quantitative RT-PCR (ND = not detected). No HPV-16 DNA was detected in oral rinse of patients with p16INK4a-negative tumors
Fig. 2
Fig. 2
HPV-16 RNA in oral rinse associates with tumor p16INK4a status. a Detection of HPV-16 RNA in representative oral rinse samples of patients with p16INK4a-positive and-negative tumors using RT-PCR. (b) The analysis of HPV-16 RNA copy number in oral rinse of patients with p16INK4a-positive tumors using quantitative RT-PCR. (ND = Not detected). No HPV-16 RNA was detected in oral rinse of patients with p16-negative tumors
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