Evaluating the Utility and Prevalence of HPV Biomarkers in Oral Rinses and Serology for HPV-related Oropharyngeal Cancer

Abstract

Performance of commercially available human papillomavirus (HPV) assays (approved for cervical HPV detection) is unknown for detecting HPV-related oropharyngeal cancer (HPV-OPC). Assays for detection of HPV DNA [ELISA (DEIA) and Cobas], and RNA (Aptima) in oral rinse samples, and serum HPV oncogene antibodies were evaluated. Sensitivity and specificity of each test was explored among HPV-OPC cases and controls. Biomarker prevalence was evaluated among 294 "at-risk" people (screening) and 133 "high-risk" people [known to previously have oral oncogenic HPV (oncHPV) DNA and/or HPV16 E6/E7 antibodies detected]. HPV16 E6 antibodies had the best overall test performance with sensitivity of 88%, compared with oral HPV16 DNA sensitivity of 51% by DEIA and 43% by Cobas (each P < 0.001). Specificity was comparable in each of these tests (!98%). When positivity for any oncHPV type was compared with HPV16 for the same test, sensitivity was comparable (60% vs. 51%, 40% vs. 43%, and 92% vs. 88% for DEIA, Cobas, and E6 antibodies, respectively), but specificity was reduced (93%-97%). Aptima had poor sensitivity (23%). Sensitivity decreased when cotesting HPV16 oral rinse DNA and E6 antibodies (37%-48%), or multiple E antibodies (69%-72%). HPV16 DNA were detected in $2% of the at-risk by either DEIA or Cobas and up to 15% of the high-risk population. HPV16 E6 seroprevalence was 2.3% and 2.4% in the at-risk and high-risk populations, respectively. Oral rinse HPV testing had moderate-to-poor sensitivity for HPV-OPC, suggesting many true positives would be missed in a potential screening scenario. HPV16 E6 serum antibody was the most promising biomarker evaluated.

Long-term Persistence of Oral HPV Over 7 Years of Follow-up

“…The biomarker used in this study—oncogenic oral HPV DNA—has moderate sensitivity but good specificity ( 14 , 28 ) for HPV-OPC, which would limit its utility (as a stand-alone test) for any screening program. Even among at-risk groups, oral HPV-16 prevalence is low, at no more than 4% ( 14 , 29 ). Although oral HPV persistence is clearly a prerequisite to development of HPV-OPC, our data suggest a large number is needed to screen to detect a single cancer (poor positive predictive value).…”

“…Oral rinse samples were collected using 10 mL saline or Scope and a 30-second oral rinse and gargle as previously described ( 14 ). Samples were stored at 4°C until processed.…”

“…Oral rinse samples were tested in the laboratory of Dr Maura Gillison at The Ohio State University (for the POPS study) and in the DDL Diagnostic Laboratory ( https://www.ddl.nl for the MOUTH study). Oral HPV DNA detection involved polymerase chain reaction (PCR) with the PGMY09/11 primer system in the Gillison Lab ( 12 ) and with the SPF10 primer system in the DDL lab (version 1, Labo Bio-medical Products, Rijswijk, the Netherlands) ( 14 ). HPV type specification was conducted using Roche linear array in the Gillison lab and the SPF10 DEIA/LIPA system in the DDL lab.…”

Development and validation of a multiplex immunoassay for the simultaneous quantification of type-specific IgG antibodies to E6/E7 oncoproteins of HPV16 and HPV18

“…While we hypothesized that HPV+ subjects would have greater antibody concentrations, we found a modest antibody concentration increase in HPV+ subjects to normal healthy donors. This may be attributed to the broad prevalence of HPV16 and 18 positivity in the general population due to the number of sexual partners [24,25] lending itself the need to identify a true negative population to set serostatus cutoff values from individuals with less than 1 sexual partner. Furthermore, we found low antibody concentrations in our pediatric donor cohort which may be attributed to material antibodies [21] as these children were between ages 1-5 days.…”

Abnormal cytology in oropharyngeal brushings and in oral rinses is not associated with HPV infection: The OHMAR study

“…Currently, the utility of HPV testing on oral samples seems to be limited, 16 whereas serologic testing for HPV-16 E6 protein appears to be the most promising tool. 17,18 However, this has not been validated, and its implementation is still a long way off. Oral cytology has also been explored, but the only attempt to evaluate the association between oral HPV infection and cytologic abnormalities of the oropharynx in a population at risk (ie, HIV-infected individuals) suggested the unfeasibility of an oral Papanicolaou (Pap)-test equivalent.…”