ABSTRACT
BACKGROUND: Determination of human papillomavirus (HPV) status has become clinically relevant for patient stratification under UICC TNM8 staging. Within the United Kingdom, a combination of p16 IHC and HPV DNA-ISH is recommended for classifying HPV status. This study will assess a series of clinically applicable second-line molecular tests to run in combination with p16 IHC to optimally determine HPV status. METHODS: The ability of HPV RNA-ISH, HPV DNA-ISH, and HPV DNA-PCR to identify p16-positive/HPV-positive patients was investigated in a population-based oropharyngeal squamous cell carcinoma (OPSCC) cohort of patients diagnosed in Northern Ireland from 2000 to 2011. RESULTS: Only 41% of the Northern Irish OPSCC patient population was associated with HPV-driven carcinogenesis. Both ISH assays were more specific than the DNA-PCR assay (100% and 95% vs. 67%) and were less likely to be affected by preanalytic factors such as increasing block age. A pooled HPV genotype probe for RNA-ISH was found to be the most accurate molecular assay assessed (95% accuracy) when compared with p16 positivity. CONCLUSIONS: Our study demonstrates the advantage of tissue-based molecular assays when determining HPV status in retrospective samples. Specifically, we demonstrate the enhanced sensitivity and specificity of ISH techniques compared with PCR-based methodology when working with formalin-fixed paraffin-embedded tissue, and found HPV RNA-ISH to be the most effective assay for determining HPV status. IMPACT: As p16 IHC is a relatively inexpensive, accessible, and sensitive test for stratifying patients by HPV status, this study finds that more patients would benefit from first-line p16 IHC followed by specific HPV testing using HPV RNA-ISH to confirm HPV status.
Subject(s)
Alphapapillomavirus/isolation & purification , Human Papillomavirus DNA Tests/methods , Oropharyngeal Neoplasms/diagnosis , Papillomavirus Infections/diagnosis , Squamous Cell Carcinoma of Head and Neck/diagnosis , Age Factors , Alphapapillomavirus/genetics , Alphapapillomavirus/immunology , Biomarkers, Tumor/analysis , Biomarkers, Tumor/immunology , Cyclin-Dependent Kinase Inhibitor p16/analysis , Cyclin-Dependent Kinase Inhibitor p16/immunology , DNA, Viral/isolation & purification , Female , Follow-Up Studies , Humans , Immunohistochemistry , In Situ Hybridization , Kaplan-Meier Estimate , Male , Middle Aged , Neoplasm Staging , Northern Ireland/epidemiology , Oropharyngeal Neoplasms/immunology , Oropharyngeal Neoplasms/pathology , Oropharyngeal Neoplasms/virology , Papillomavirus Infections/mortality , Papillomavirus Infections/pathology , Papillomavirus Infections/virology , Prognosis , RNA, Viral/isolation & purification , Retrospective Studies , Sensitivity and Specificity , Squamous Cell Carcinoma of Head and Neck/mortality , Squamous Cell Carcinoma of Head and Neck/pathology , Squamous Cell Carcinoma of Head and Neck/virologyABSTRACT
BACKGROUND: TNM8 staging for oropharyngeal squamous cell carcinomas (OPSCC) surrogates p16 immunohistochemistry for HPV testing. Patients with p16+ OPSCC may lack HPV aetiology. Here, we evaluate the suitability of TNM8 staging for guiding prognosis in such patients. METHODS: HPV status was ascertained using p16 immunohistochemistry and high-risk HPV RNA and DNA in situ hybridisation. Survival by stage in a cohort of OPSCC patients was evaluated using TNM7/TNM8 staging. Survival of p16+/HPV- patients was compared to p16 status. RESULTS: TNM8 staging was found to improve on TNM7 (log rank p = 0·0190 for TNM8 compared with p = 0·0530 for TNM7) in p16+ patients. Patients who tested p16+ but were HPV- (n = 20) had significantly reduced five-year survival (33%) compared to p16+ patients (77%) but not p16- patients (35%). Cancer stage was reduced in 95% of p16+/HPV- patients despite having a mortality rate twice (HR 2.66 [95% CI: 1.37-5.15]) that of p16+/HPV+ patients under new TNM8 staging criteria. CONCLUSION: Given the significantly poorer survival of p16+/HPV- OPSCCs, these data provide compelling evidence for use of an HPV-specific test for staging classification. This has particular relevance in light of potential treatment de-escalation that could expose these patients to inappropriately reduced treatment intensity as treatment algorithms evolve.
Subject(s)
Oropharyngeal Neoplasms/genetics , Papillomavirus Infections/genetics , Viral Proteins/genetics , Adolescent , Adult , Child , Child, Preschool , Disease-Free Survival , Female , Gene Expression Regulation, Neoplastic/genetics , Humans , Infant , Infant, Newborn , Male , Middle Aged , Neoplasm Staging , Oropharyngeal Neoplasms/mortality , Oropharyngeal Neoplasms/pathology , Oropharyngeal Neoplasms/virology , Papillomavirus Infections/pathology , Papillomavirus Infections/virology , Young AdultABSTRACT
For a number of years, scientists have been trying to develop aids that can make visually impaired people more independent and aware of their surroundings. Computer-based automatic navigation tools are one example of this, motivated by the increasing miniaturization of electronics and the improvement in processing power and sensing capabilities. This paper presents a complete navigation system based on low cost and physically unobtrusive sensors such as a camera and an infrared sensor. The system is based around corners and depth values from Kinect's infrared sensor. Obstacles are found in images from a camera using corner detection, while input from the depth sensor provides the corresponding distance. The combination is both efficient and robust. The system not only identifies hurdles but also suggests a safe path (if available) to the left or right side and tells the user to stop, move left, or move right. The system has been tested in real time by both blindfolded and blind people at different indoor and outdoor locations, demonstrating that it operates adequately.
