Jesper Bonde1, Ditte Møller Ejegod2, Kate Cuschieri3, Joakim Dillner4, Daniëlle A M Heideman5, Wim Quint6, Miguel Angel Pavon Ribas7, Elizaveta Padalko8, Irene Kraus Christiansen9, Lan Xu10, Marc Arbyn10. 1. Molecular Pathology Laboratory, Department of Pathology, Copenhagen University Hospital, Kettegårds Alle 30, 2650 Hvidovre, Denmark. Electronic address: jesper.hansen.bonde@regionh.dk. 2. Molecular Pathology Laboratory, Department of Pathology, Copenhagen University Hospital, Kettegårds Alle 30, 2650 Hvidovre, Denmark. 3. Scottish HPV Reference Laboratory, Royal Infirmary of Edinburgh, EH16 4SA Edinburgh, Scotland, United Kingdom. 4. International HPV Reference Center, Karolinska University Hospital, Alfred Nobels Allê 8, Huddinge, Stockholm, Sweden. 5. Cancer Center Amsterdam, Department of Pathology, VU University Medical Center, De Boelelaan 1118, 1081 HV Amsterdam, The Netherlands. 6. DLL Diagnostic Laboratory, Visseringlaan 25, 2288 ER Rijswijk, The Netherlands. 7. Infection and Cancer Laboratory, Cancer Epidemiology Research Program and CIBER-ONC, Granvia de L'Hospitalet 199-203, Institut Català d'Oncologia, Barcelona, Spain. 8. Department of Laboratory Medicine, Ghent University Hospital, C. Heymanslaan 10, B-9000 Ghent, Belgium. 9. Norwegian HPV Reference Laboratory, Department of Microbiology and Infection Control, Akershus University Hospital, 1478 Lørenskog, Norway. 10. Sciensano, Belgian Cancer Centre/Unit of Cancer Epidemiology Juliette Wytsmanstreet 14, B1050 Brussels, Belgium.
Abstract
BACKGROUND: The VALidation of HPV GENoyping Tests (VALGENT) is an international initiative designed to validate HPV assays with genotyping capability. The VALGENT4 protocol differs from previous VALGENT installments as the sample collection medium is SurePath, and exclusively includes samples from women ≥30 years of age which is concordant with the majority of HPV primary screening guidelines. Here we present the protocol for the fourth installment of the VALGENT framework. OBJECTIVES: In VALGENT4 11 HPV assays will be evaluated using two comparator assays based on PCR with the GP5+/6+ primers. STUDY DESIGN: Overall, the VALGENT4 panel consists of 1,297 routine samples comprised of 998 unselected, consecutive samples, of which 51 samples had abnormal cytology with 13 women diagnosed with ≥CIN2, and 299 consecutive samples enriched for ≥ASCUS cytology (100 ASCUS, 100 LSIL, 99 HSIL) with 106 ≥CIN2 upon follow up. Manipulated and DNA extracted panel samples were characterized with respect to human beta globin (HBB) and overall DNA content and composition to quality assess the panel prior to distribution to the collaborating sites. RESULT: The relative cellularity (mean CT value of HBB from the Onclarity assay) on the 1,297 LBC samples (CT=24.8) was compared with 293 un-manipulated routine cytology screening samples (CT=23.8). Furthermore, the DNA extracted panel samples was characterized using the Exome iPLEX pro assay, which reports amplifiable copies on individual samples as well as copies of five different base pair lengths. Here the data showed a slightly lower number of amplifiable DNA copies (ratio: 0.7, p=<0.01)) in the VALGENT4 panel samples compared to routine extracted cervical DNA samples CONCLUSION: The present manuscript details the manipulation, processing and quality assessment of samples used in VALGENT-4. This methodological document may be of value for future international projects of HPV test validation.
BACKGROUND: The VALidation of HPV GENoyping Tests (VALGENT) is an international initiative designed to validate HPV assays with genotyping capability. The VALGENT4 protocol differs from previous VALGENT installments as the sample collection medium is SurePath, and exclusively includes samples from women ≥30 years of age which is concordant with the majority of HPV primary screening guidelines. Here we present the protocol for the fourth installment of the VALGENT framework. OBJECTIVES: In VALGENT4 11 HPV assays will be evaluated using two comparator assays based on PCR with the GP5+/6+ primers. STUDY DESIGN: Overall, the VALGENT4 panel consists of 1,297 routine samples comprised of 998 unselected, consecutive samples, of which 51 samples had abnormal cytology with 13 women diagnosed with ≥CIN2, and 299 consecutive samples enriched for ≥ASCUS cytology (100 ASCUS, 100 LSIL, 99 HSIL) with 106 ≥CIN2 upon follow up. Manipulated and DNA extracted panel samples were characterized with respect to humanbeta globin (HBB) and overall DNA content and composition to quality assess the panel prior to distribution to the collaborating sites. RESULT: The relative cellularity (mean CT value of HBB from the Onclarity assay) on the 1,297 LBC samples (CT=24.8) was compared with 293 un-manipulated routine cytology screening samples (CT=23.8). Furthermore, the DNA extracted panel samples was characterized using the Exome iPLEX pro assay, which reports amplifiable copies on individual samples as well as copies of five different base pair lengths. Here the data showed a slightly lower number of amplifiable DNA copies (ratio: 0.7, p=<0.01)) in the VALGENT4 panel samples compared to routine extracted cervical DNA samples CONCLUSION: The present manuscript details the manipulation, processing and quality assessment of samples used in VALGENT-4. This methodological document may be of value for future international projects of HPV test validation.
Authors: Marc Arbyn; Sharonjit Kaur Dhillon; Marianna Martinelli; Cindy Simoens; Lize Cuypers; Jannes Bode; Marc Van Ranst; Philippe Corbisier; Jesper Bonde; Clementina Cocuzza Journal: Arch Public Health Date: 2022-03-29
Authors: Lindsey Asti; Colin Hopley; Cameron Avelis; Sarah M Bartsch; Leslie E Mueller; Molly Domino; Sarah N Cox; Jeffrey C Andrews; Samuel L Randall; Owen J Stokes-Cawley; Caitlin Asjes; Bruce Y Lee Journal: Sex Transm Dis Date: 2021-05-01 Impact factor: 2.830
Authors: Jesper Bonde; Arno Floore; Ditte Ejegod; Frederique J Vink; Albertus Hesselink; Peter M van de Ven; Anja Oštrbenk Valenčak; Helle Pedersen; Saskia Doorn; Wim G Quint; Karl Ulrich Petry; Mario Poljak; Grazyna Stanczuk; Kate Cuschieri; Silvia de Sanjosé; Maaike Bleeker; Johannes Berkhof; Chris J L M Meijer; Daniëlle A M Heideman Journal: Int J Cancer Date: 2020-10-21 Impact factor: 7.396