National Reference Center (NRC) for Human papillomavirus

Presentation of the NRC

The species of human papillomaviruses (HPV) comprises more than 200 genotypes among which a dozen are classified by the International Agency for Research in Cancer as carcinogenic. Persistent infection with a carcinogenic (often called high-risk) HPV type is necessary but not sufficient for developing cervical cancer. HPV can also cause other anogenital cancers or oropharyngeal cancers. Cervical cancer is a major public health problem. In 2020, more than 604,000 new cases and 341,000 deaths were reported worldwide (Singh et al., Lancet Glob Health 2023⁵). This renders cervical cancer the third in the World.

The National Reference Center (NRC) for Human Papillomavirus (HPV) is a consortium of three different institutes. Sciensano, Algemeen Medisch Laboratorium (AML) and the University Hospital of Ghent (UZGent) bring their own expertise for quality assurance of HPV diagnosis, for proposing HPV proficiency panels, for HPV surveillance and other HPV projects related to the activities of the NRC.

Sciensano has a world-widely recognized expertise in HPV test validation whereas AML has an extensive experience in routine HPV testing with the use of the in-house, automated and clinically validated RIATOL HPV genotyping qPCR (Dhillon et al., J Med 2023³; Benoy et al., J Clin Virol 2019⁶). High volumes of samples can be run in a short timeframe. UZGent contributes to the NRC by offering their expertise Next Generation Sequencing (NGS) using an in-house targeted NGS assay. Request forms for HPV testing for research or monitoring purposes are available by clicking on the links on the panel displayed on the right side of the screen.

The NRC for HPV is involved in several projects such as:

• The development of a proficiency test with the collaboration of Quality of Laboratories (Sciensano) for assessing HPV-based screening assays. On a yearly basis, a panel of samples is sent to first-line laboratories.
• The surveillance of HPV genotypes in Belgium. On a yearly basis, screening samples from the three regions are collected and analysed for HPV. A map of HPV genotypes is drawn and the eventual risk of drift towards genotypes not targeted by the commercialized vaccines is assessed.
• A standard operating procedure for promoting quality in HPV testing by re-analysing seemingly HPV negative high grade squamous intraepithelial lesions (HSIL) and invasive cervical cancer was recently proposed together with the International HPV Reference Center (Pretét et al., Preprint⁷). It involves the referral of HPV-negative HSIL and invasive cervical cancer to the NRC for retesting and eventual subsequent NGS assay for confirmation of HPV negativity or subsequent identification and characterization of HPV.

Available tests

List of assays performed by the NRC for HPV :

• RIATOL HPV genotyping qPCR.
  The RIATOL HPV genotyping qPCR assay was developed at AML, Sonic Healthcare Benelux (Antwerp, Belgium) and used in routine screening for more than 15 years under ISO15189 accreditation. The assay comprises a multiplex RT-PCR that detects viral HPV DNA in a quantitative way. RIATOL qPCR is capable of detecting E6 or E7 genes of 18 distinct HPV types separately (i.e. HPV6, 11, 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 56, 58, 59, 66, 67, and 68). In addition, human βeta-globin is used as both an endogenous cellular quality control and as relative normalizer of viral copies detected for HPV-positive samples.
• HPV targeted in-house NGS.