Recent improvements in in situ hybridization for the detection of HPV infections in clinical samples

WCRJ 2020; 7: e1542
DOI: 10.32113/wcrj_20203_1542

  Topic: Gynaecological cancer, Virus and cancer     Category:

Abstract

Objective: Human Papilloma Virus (HPV) infection is well-established as a cause of cervical cancer. Importantly, early HPV detection can decrease both the frequency and mortality of HPV-related cancers. In situ hybridization (ISH) is a widely used method for the early detection of HPV. Yet, ISH can be expensive, time-consuming and, in some cases, insufficiently sensitive to detect nucleic acid target at low copy number, which may lead to false-positive or false-negative results. To address these limitations, we recently developed a novel in situ hybridization technology based on proprietary Loop RNA probes (LRPs), which provides enhanced sensitivity, high-specificity and improved cost-effectiveness.

Patients and Methods: Manual and automated ISH was performed on paraffin-embedded cervical cancer cell lines and cervical biopsy tissues obtained from HPV-positive and -negative patients. ISH was also performed on liquid-based cytology samples, spread in monolayer, for the detection of HPV in cervicovaginal samples.

Results: We compared our Loop RNA probes and reagents with commercially available kits for detecting HPV. LRPs were able to detect a single copy genome- integrated HPV, as well as HPV RNA in cell lines, patient biopsies and in liquid-based cytology samples.

Conclusions: Our results show that LRP technology is a powerful system for the in-situ detection of HPV DNA and RNA at low copy number, even down to a single copy of genome-integrated HPV.

To cite this article

Recent improvements in in situ hybridization for the detection of HPV infections in clinical samples

WCRJ 2020; 7: e1542
DOI: 10.32113/wcrj_20203_1542

Publication History

Submission date: 11 Feb 2020

Revised on: 03 Mar 2020

Accepted on: 05 Mar 2020

Published online: 12 Mar 2020