Developing a new diagnostic algorithm for human papilloma virus associated oropharyngeal carcinoma: An investigation of HPV DNA assays

Abstract

BackgroundHuman papilloma virus (HPV) has been implicated in the development of a large proportion of oropharyngeal squamous cell carcinoma (OPSCC). Current techniques used to diagnose HPV etiology require histopathologic analysis. We aim to investigate the diagnostic accuracy of a new application non-histopathologic diagnostic tests to help assist diagnosis of HPV-related oropharyngeal tumors.MethodsPatients with OPSCC with nodal metastasis were consecutively recruited from a multidisciplinary cancer clinic. Appropriate samples were collected and analyzed. The various tests examined included COBAS® 4800, Cervista® HR and Genotyping. These tests were compared to p16 staining, which was used as the diagnostic standard. StataIC 14.2 was used to perform analysis, including sensitivity, specificity and receiver operator characteristic [ROC] curves.ResultsThe COBAS® FNA (area under ROC 0.863) and saliva (area under ROC 0.847) samples performed well in diagnosing HPV positive and negative tumors. Samples tested with Cervista® did not corroborate p16 status reliably. We were able to increase the diagnostic yield of the COBAS® FNA samples by applying the results of the saliva test to negative FNA samples which correctly identified 11 additional p16 positive tumors (area under ROC 0.915).ConclusionSurrogate testing for HPV using alternate methods is feasible and closely predicts the results of standard diagnostic methods. In the future, these could minimize invasive procedures for diagnosing HPV-related oropharyngeal cancer, but also help to diagnose and treat patients with unknown primaries.

Exposure to Microbial Metabolite Butyrate Prolongs the Survival Time and Changes the Growth Pattern of Human Papillomavirus 16 E6/E7-Immortalized Keratinocytes in Vivo

“…OPSCC is known to develop cervical nodal metastases in the absence of morphologic features of conventional invasion, and in-situ diagnosis is not used at this anatomical site for practical purposes. The lesions are often small and clinically occult, and patients frequently present with lymph node metastases as compared with HPV negative tumors that commonly present with pain 57 . It is generally accepted that the lack of a continuous basement membrane of the reticular epithelium provides a weak barrier, allowing the HPV-infected cells to develop an invasive tumor front by budding off the basal cell layer with lesser restraint.…”

“…The lesions are often small and clinically occult, and patients frequently present with lymph node metastases compared with HPV-negative tumors that commonly present with pain. 58 It is generally accepted that the lack of a continuous basement membrane of the reticular epithelium provides a weak barrier, allowing the HPV-infected cells to develop an invasive tumor front by budding off the basal cell layer with lesser restraint. In this study, MOKE6E7 cells showed no organized basement membrane in vivo , but they still formed epithelial lesions confluent with the keratinocytes around the main lumens, likened to the precursor lesions in the conventional SCC.…”

Current salivary biomarkers for detection of human papilloma virus‐induced oropharyngeal squamous cell carcinoma

“…These statistics were much lower than their fine needle aspirate counterparts. Similar analysis was also done by the same group to evaluate Hologic's Cervista HR and Cervista HPV Genotype (detects specific DNA sequences and targets HPV E6/E7 oncogenes, Marlborough, MA) 104 . Oropharyngeal swabs from 37 patients resulted in a sensitivity of 100% for both tests, with specificities of 33% and 14%, respectively.…”

“…Similar analysis was also done by the same group to evaluate Hologic's Cervista HR and Cervista HPV Genotype (detects specific DNA sequences and targets HPV E6/E7 oncogenes, Marlborough, MA). 104 Oropharyngeal swabs from 37 patients resulted in a sensitivity of 100% for both tests, with specificities of 33% and 14%, respectively. A similar group recently assessed Atila BioSystems' Ampfire HPV assay (isothermal PCR identification of "various regions in the viral genome including E6/E7," Mountain View, CA); oropharyngeal swabs and saliva resulted in a reported sensitivity of 40.3% and a specificity of 93.3% among 77 tumors.…”

An algorithm to evaluate the efficacy of detecting somatic mutations

“…Somatic mutations are tested for in a variety of heterogeneous cell sample types, including solid tumors, liquid biopsy, mammalian cell culture, stool, urine, saliva, cerebrospinal fluids, lung lavage, gastric, and tissue and/or organ transplants. [1][2][3][4][5][6][7][8][9] Further, the genetic variants could be single nucleotide polymorphism, multiple mutations, deletion, addition of nucleotides, epigenetic changes (e.g. Methylation), gene expression, splice variants and copy number variations of all the above genetic variants.…”