Abstract
BackgroundHigh-risk human papillomavirus (HR-HPV) is notoriously associated with tumor progression in a broad spectrum of malignancies. Detection of HR-HPV is clinically important in the management of HPV-related carcinomas, particularly in cervical cancer and oropharyngeal squamous cell carcinoma (OPSCC). Several methods for HPV detection are currently available including Polymerase chain reaction (PCR)-based techniques, DNA in situ hybridization (ISH), RNA ISH, and p16 immunohistochemistry (IHC). Currently, the guidelines for HPV detection in cervical carcinoma are available, while no clear consensus has not yet been reached on the gold standard for HPV testing in OPSCC. Multimodality testing could help to reliably identify patients with transcriptionally active high-risk HPV-positive.MethodsWe propose a multiplex approach carrying out HPV RNA ISH and p16 IHC on the same slide to detect simultaneously HPV E6/E7 transcripts and p16INK4a overexpression. We tested this assay in two different series one of the cervical cancers with p16-positive, as control, and the other of oropharyngeal squamous cell carcinomas with blind p16 status.ResultsThe multiplex HPV RNA ISH /p16 IHC results in the series both of the cervical cancers and the oral-oropharyngeal cancers were fully concordant with the previous results achieved through the classic p16 IHC and HPV RNA scope carried out on two different slides.ConclusionsOur results suggesting several advantages of this technical approach, namely an easy interpretation fully in the light field, the feasibility in formalin-fixed paraffin-embedded tissue sections, complete automation and a potential wide spreadable for routine testing in several clinical laboratories.
Highlights
High-risk human papillomavirus (HR-Human papillomavirus (HPV)) is notoriously associated with tumor progression in a broad spectrum of malignancies
The new guidelines proposed by the College of American Pathologists (CAP), for HPV testing in head and neck carcinomas in routine clinical practice proposed the interpretation of p16 IHC as follows: tumors with lack of staining or < 70% of nuclear and cytoplasmic staining are classified as HPV negative, while tumors with ≥70% of nuclear and cytoplasmic staining as HPV positive [10]
Results in the series of cervical cancers The cervical cancers selected from our archive on the basis of p16-positive IHC status were tested for RNAscope HPV-test
Summary
High-risk human papillomavirus (HR-HPV) is notoriously associated with tumor progression in a broad spectrum of malignancies. Detection of HR-HPV is clinically important in the management of HPV-related carcinomas, in cervical cancer and oropharyngeal squamous cell carcinoma (OPSCC). The guidelines for HPV detection in cervical carcinoma are available, while no clear consensus has not yet been reached on the gold standard for HPV testing in OPSCC. 200 different HPV genotypes have been identified responsible of a broad spectrum of the clinical profiles, from benign lesions to HPV-related carcinomas [1, 2]. The European guidelines for quality assurance in cervical cancer screening recommend to test for HPV alone in women aged 30–65 every five years [8]
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