Human papillomavirus negative high grade cervical lesions and cancers: Suggested guidance for HPV testing quality assurance

Jean Luc Prétet¹, Laila Sara Arroyo Mühr², Kate Cuschieri³, María Dolores Fellner⁴, Rita Mariel Correa⁴, María Alejandra Picconi⁴, Suzanne M Garland⁵, Gerald L Murray⁵, Monica Molano⁶, Michael Peeters⁷, Steven Van Gucht⁷, Charlotte Lambrecht⁸, Davy Vanden Broeck⁸, Elizaveta Padalko⁹, Marc Arbyn¹⁰, Quentin Lepiller¹, Alice Brunier¹, Steffi Silling¹¹, Kristiane Søreng¹², Irene Kraus Christiansen¹², Mario Poljak¹³, Camilla Lagheden², Emel Yilmaz², Carina Eklund², Hem R Thapa¹⁴, Troy D Querec¹⁴, Elizabeth R Unger¹⁴, Joakim Dillner¹⁵

Affiliations

¹ French National Papillomavirus Reference Center, CHU de Besançon, EA3181, Université of Franche-Comté, F-25000 Besançon, France.
² Swedish National HPV Reference Laboratory, Center for Cervical Cancer Elimination, Karolinska Institutet and Karolinska University Hospital F56, 141 86 Stockholm, Sweden.
³ Scottish HPV Reference Laboratory, Deptartment of Lab Medicine, Royal Infirmary of Edinburgh, Edinburgh, EH16 4SA, UK.
⁴ Laboratorio Nacional y Regional de Referencia de Papilomavirus Humanos, Instituto Nacional de Enfermedades Infecciosas-ANLIS "Dr Malbrán", C1282AFF Buenos Aires, Argentina.
⁵ Royal Women's Hospital, Parkville 3052 VIC, Australia; Department of Obstetrics and Gynaecology, University of Melbourne, Parkville 3052 VIC, Australia.
⁶ Royal Women's Hospital, Parkville 3052 VIC, Australia.
⁷ National Reference Centre for Human Papillomaviruses, Viral Diseases, Infectious Diseases in Humans, Sciensano. Rue Juliette Wytsman 14, 1050 Brussels, Belgium.
⁸ National Reference Centre for Human Papillomaviruses, Algemeen Medisch Laboratorium. Emiel Vloorsstraat 9, 2020 Antwerp, Belgium.
⁹ National Reference Centre for Human Papillomaviruses, Laboratory of Medical Microbiology, University Hospital Ghent. C. Heymanslaan 10, 9000 Ghent, Belgium.
¹⁰ Unit Cancer Epidemiology, Cancer centre, Sciensano. Rue Juliette Wytsman 14, 1050 Brussels, Belgium.
¹¹ Institute of Virology, National Reference Center for Papilloma- and Polyomaviruses, University of Cologne, Faculty of Medicine and University Hospital of Cologne, 50935 Cologne, Germany.
¹² Norwegian HPV Reference Laboratory, Department of Microbiology and Infection Control, Akershus University Hospital, Lørenskog, Norway.
¹³ Institute of Microbiology and Immunology, Faculty of Medicine, University of Ljubljana, Zaloška 4, 1000 Ljubljana, Slovenia.
¹⁴ Division of High-Consequence Pathogens and Pathology, Centers for Disease Control and Prevention, Atlanta, GA 30329, USA.
¹⁵ Swedish National HPV Reference Laboratory, Center for Cervical Cancer Elimination, Karolinska Institutet and Karolinska University Hospital F56, 141 86 Stockholm, Sweden. Electronic address: Joakim.dillner@ki.se.

PMID: 38401369
PMCID: PMC11863830
DOI: 10.1016/j.jcv.2024.105657

Human papillomavirus negative high grade cervical lesions and cancers: Suggested guidance for HPV testing quality assurance

Jean Luc Prétet et al. J Clin Virol. 2024 Apr.

. 2024 Apr:171:105657.

doi: 10.1016/j.jcv.2024.105657. Epub 2024 Feb 20.

Authors

Affiliations

¹ French National Papillomavirus Reference Center, CHU de Besançon, EA3181, Université of Franche-Comté, F-25000 Besançon, France.
² Swedish National HPV Reference Laboratory, Center for Cervical Cancer Elimination, Karolinska Institutet and Karolinska University Hospital F56, 141 86 Stockholm, Sweden.
³ Scottish HPV Reference Laboratory, Deptartment of Lab Medicine, Royal Infirmary of Edinburgh, Edinburgh, EH16 4SA, UK.
⁴ Laboratorio Nacional y Regional de Referencia de Papilomavirus Humanos, Instituto Nacional de Enfermedades Infecciosas-ANLIS "Dr Malbrán", C1282AFF Buenos Aires, Argentina.
⁵ Royal Women's Hospital, Parkville 3052 VIC, Australia; Department of Obstetrics and Gynaecology, University of Melbourne, Parkville 3052 VIC, Australia.
⁶ Royal Women's Hospital, Parkville 3052 VIC, Australia.
⁷ National Reference Centre for Human Papillomaviruses, Viral Diseases, Infectious Diseases in Humans, Sciensano. Rue Juliette Wytsman 14, 1050 Brussels, Belgium.
⁸ National Reference Centre for Human Papillomaviruses, Algemeen Medisch Laboratorium. Emiel Vloorsstraat 9, 2020 Antwerp, Belgium.
⁹ National Reference Centre for Human Papillomaviruses, Laboratory of Medical Microbiology, University Hospital Ghent. C. Heymanslaan 10, 9000 Ghent, Belgium.
¹⁰ Unit Cancer Epidemiology, Cancer centre, Sciensano. Rue Juliette Wytsman 14, 1050 Brussels, Belgium.
¹¹ Institute of Virology, National Reference Center for Papilloma- and Polyomaviruses, University of Cologne, Faculty of Medicine and University Hospital of Cologne, 50935 Cologne, Germany.
¹² Norwegian HPV Reference Laboratory, Department of Microbiology and Infection Control, Akershus University Hospital, Lørenskog, Norway.
¹³ Institute of Microbiology and Immunology, Faculty of Medicine, University of Ljubljana, Zaloška 4, 1000 Ljubljana, Slovenia.
¹⁴ Division of High-Consequence Pathogens and Pathology, Centers for Disease Control and Prevention, Atlanta, GA 30329, USA.
¹⁵ Swedish National HPV Reference Laboratory, Center for Cervical Cancer Elimination, Karolinska Institutet and Karolinska University Hospital F56, 141 86 Stockholm, Sweden. Electronic address: Joakim.dillner@ki.se.

PMID: 38401369
PMCID: PMC11863830
DOI: 10.1016/j.jcv.2024.105657

Abstract

Background: Some high-grade cervical lesions and cervical cancers (HSIL+) test negative for human papillomavirus (HPV). The HPV-negative fraction varies between 0.03 % and 15 % between different laboratories. Monitoring and extended re-analysis of HPV-negative HSIL+ could thus be helpful to monitor performance of HPV testing services. We aimed to a) provide a real-life example of a quality assurance (QA) program based on re-analysis of HPV-negative HSIL+ and b) develop international guidance for QA of HPV testing services based on standardized identification of apparently HPV-negative HSIL+ and extended re-analysis, either by the primary laboratory or by a national HPV reference laboratory (NRL).

Methods: There were 116 initially HPV-negative cervical specimens (31 histopathology specimens and 85 liquid-based cytology samples) sent to the Swedish HPV Reference Laboratory for re-testing. Based on the results, an international QA guidance was developed through an iterative consensus process.

Result: Standard PCR testing detected HPV in 55.2 % (64/116) of initially "HPV-negative" samples. Whole genome sequencing of PCR-negative samples identified HPV in an additional 7 samples (overall 61.2 % HPV positivity). Reasons for failure to detect HPV in an HSIL+ lesion are listed and guidance to identify cases for extended re-testing, including which information should be included when referring samples to an NRL are presented.

Conclusion: Monitoring the proportion of and reasons for failure to detect HPV in HSIL+ will help support high performance and quality improvement of HPV testing services. We encourage implementation of QA strategies based on re-analysis of "HPV negative" HSIL+ samples.

Keywords: Cervical cancer; HPV testing; HSIL; Human papillomavirus; Quality assurance; Screening.

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Conflict of interest statement

Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: K Cuschieri's institution has received research funding or gratis consumables to support research from the following commercial entities in the last 3 years: Cepheid, Euroimmun, GeneFirst, SelfScreen, Hiantis, Seegene, Roche, Abbott, Hologic and Vaccitech. M.P.’s institution received research funding, free-of-charge reagents, and consumables to support research in the last 3 years from Qiagen, Seegene, Abbott, and Roche, all paid to his employer. Sciensano the employer of M.A. received funding in the framework of Valgent and VALHUDES, which are 2 researcher induced protocols for evaluation of HPV tests on cervical and vaginal samples respectively (see Arbyn et J Clin Virol 2016 & 2018). M.A, did not receive any financial or material benefit from these projects.

Figures

**Figure 1:**
Flowchart for HPV confirmatory testing

See this image and copyright information in PMC

References

1. Walboomers JM, Jacobs MV, Manos MM, Bosch FX, Kummer JA, Shah KV, Snijders PJ, Peto J, Meijer CJ, Munoz N. Human papillomavirus is a necessary cause of invasive cervical cancer worldwide. J Pathol 1999;189: 12–9. - PubMed
1. Arroyo Muhr LS, Lagheden C, Lei J, Eklund C, Nordqvist Kleppe S, Sparen P, Sundstrom K, Dillner J. Deep sequencing detects human papillomavirus (HPV) in cervical cancers negative for HPV by PCR. Br J Cancer 2020;123: 1790–5. - PMC - PubMed
1. Cancer Genome Atlas Research N, Albert Einstein College of M, Analytical Biological S, Barretos Cancer H, Baylor College of M, Beckman Research Institute of City of H, Buck Institute for Research on A, Canada’s Michael Smith Genome Sciences C, Harvard Medical S, Helen FGCC, Research Institute at Christiana Care Health S, HudsonAlpha Institute for B, et al. Integrated genomic and molecular characterization of cervical cancer. Nature 2017;543: 378–84. - PMC - PubMed
1. Fernandes A, Viveros-Carreno D, Hoegl J, Avila M, Pareja R. Human papillomavirus-independent cervical cancer. Int J Gynecol Cancer 2022;32: 1–7. - PubMed
1. Petry KU, Cox JT, Johnson K, Quint W, Ridder R, Sideri M, Wright TC Jr., Behrens CM. Evaluating HPV-negative CIN2+ in the ATHENA trial. Int J Cancer 2016;138: 2932–9. - PMC - PubMed

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Human papillomavirus negative high grade cervical lesions and cancers: Suggested guidance for HPV testing quality assurance

Affiliations

Human papillomavirus negative high grade cervical lesions and cancers: Suggested guidance for HPV testing quality assurance

Authors

Affiliations

Abstract

Conflict of interest statement

Figures

References

Publication types

MeSH terms

Grants and funding

LinkOut - more resources

Full Text Sources

Medical