Study to investigate the prevalence of human papillomavirus in Barrett's oesophagus using a novel screening methodology

Abstract

Introduction: Human papillomavirus (HPV) is strongly associated with Barrett's dysplasia and oesophageal cancer suggesting a role in carcinogenesis. HPV persistence predicts treatment failure after endotherapy for Barrett's dysplasia. This pilot study applies a novel HPV screening tool (previously only used in the oropharynx) to detect HPV DNA directly and determine the prevalence rates in Barrett's oesophagus (BO).

Method: DNA was extracted from 20 formalin-fixed BO samples. HPV DNA was detected using real-time PCR and gel electrophoresis.

Results: 5 out of 20 patients were identified as positive for HPV. Prevalence was 25% in patients with BO.

Conclusion: This method can be used in BO's tissue to determine HPV infection. Adoption of this as a screening test could potentially revolutionise future research in this area. If a clear link between HPV and Barrett's dysplasia can be confirmed, this qPCR method has the potential to aid in monitoring and/or dysplasia detection by stratifying those most at risk and aid in the development of new therapies.

Keywords: Barrett's oesophagus; oesophageal cancer; polymerase chain reaction.

Figure 1

Laboratory L1 HPV DNA screening of archival Barrett’s oesophagus (BO) samples using Knight et al’s novel HPV screening methodology. (A) Patients 12 and 13 have a high ‘relative fluorescence’ L1 HPV DNA peak (highlighted with green arrow), indicating an abundant HPV infection, while patient 4 is negative for HPV, as indicated by no relative fluorescence L1 HPV peak (highlighted with red arrow). Patients 3, 5 and 15 have a reproducible detectable relative fluorescence peak (yellow arrow), though at lower level than patients 12 and 13, indicating a less abundant HPV infection. (B) Corresponding DNA electrophoresis showing the visualisation of L1 PCR bands, with an intense PCR band for the positive control cell line (H) and varying intensity of PCR bands for the five HPV-positive patient samples. The PCR band intensity correlates with the peak height of the relative fluorescence. HPV, human papillomavirus.