Abstract B074: Development of genetic assays for the surveillance of HPV 90 in New Orleans

Abstract

Abstract Human papillomavirus (HPV) is the main etiologic agent in 90% of anogenital and 70% of oropharyngeal cancers. Seventy percent of those cancers stem from HPV 16 or HPV 18 infection, with the remaining cases attributed to other high-risk (HR) HPV genotypes. The development and use of HPV vaccines such as Gardasil have directly affected the prevalence of HR HPVs in patient populations. The decline of dominant oncogenic strains may create an ecological vacuum, potentially allowing nondominant low-risk or unknown-risk genotypes to spread and evolve. HPV 90 was recently classified as an unknown etiologic risk and has previously demonstrated an ability to become oncogenic from a single base pair mutation within the E6 viral gene. HPV 90 has also been seen to exhibit higher prevalence than other unknown risk HPVs in patient populations similar to those populations found in New Orleans. To allow for the real-time surveillance of HPV90 during the post-vaccine era, it is our goal to establish nucleic acid-based assays that allow for sensitive and specific identification of HPV90 genes within a patient sample. The assays utilized for the surveillance of HPV90 are based on both traditional and qualitative polymerase chain reaction (PCR) technology done with HPV90 genotype specific primers for the L1 and E6 genes. The amplification of a 155bp segment of HPV90 E6 was successfully done through touchdown qPCR. The amplification of HPV L1 was done with primers and cycle methods derived from the previously published PGMY09/11 primer set. Amplicons from each PCR method were visualized via gel electrophoresis. The L1 patient amplicon will be denatured and fixed to a nylon membrane to allow for analysis by dot blot using novel HPV 90 L1 specific biotinylated probes. The patient samples utilized in this study were collected from a longitudinal study examining the genotypes found in HIV+ women in the greater New Orleans area. The cohort of HIV+ women in New Orleans (n=100) had a HPV90 prevalence of 9% when examining L1 alone and a 10% prevalence when examining E6 alone. L1 and E6 have a preliminary correlation of 90%. Testing the remaining samples from the longitudinal study will determine the persistence, prevalence and oncogenic contribution of HPV90 to patients in New Orleans. Future directions include confirming our findings utilizing our conceived dot blot assay. Once established, our L1 technique could be integrated into the clinically available Roche Linear Array genotyping assay for the additional surveillance of locally relevant HPV genotypes. Note: This abstract was not presented at the conference. Citation Format: LaKia M. Williams, Alex Berry, Jennifer Cameron. Development of genetic assays for the surveillance of HPV 90 in New Orleans [abstract]. In: Proceedings of the Eleventh AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2018 Nov 2-5; New Orleans, LA. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2020;29(6 Suppl):Abstract nr B074.