The Valgent4 protocol: Robust analytical and clinical validation of 11 HPV assays with genotyping on cervical samples collected in SurePath medium

Jesper Bonde, Ditte Møller Ejegod, Kate Cuschieri, Joakim Dillner, Daniëlle A M Heideman, Wim Quint, Miguel Angel Pavon Ribas, Elizaveta Padalko, Irene Kraus Christiansen, Lan Xu, Marc Arbyn

Research output: Contribution to journalArticleAcademicpeer-review

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.

Original languageEnglish
Pages (from-to)64-71
Number of pages8
JournalJournal of Clinical Virology
Volume108
Early online date17 Sep 2018
DOIs
Publication statusPublished - 1 Nov 2018

Cite this

Bonde, Jesper ; Ejegod, Ditte Møller ; Cuschieri, Kate ; Dillner, Joakim ; Heideman, Daniëlle A M ; Quint, Wim ; Pavon Ribas, Miguel Angel ; Padalko, Elizaveta ; Christiansen, Irene Kraus ; Xu, Lan ; Arbyn, Marc. / The Valgent4 protocol : Robust analytical and clinical validation of 11 HPV assays with genotyping on cervical samples collected in SurePath medium. In: Journal of Clinical Virology. 2018 ; Vol. 108. pp. 64-71.
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title = "The Valgent4 protocol: Robust analytical and clinical validation of 11 HPV assays with genotyping on cervical samples collected in SurePath medium",
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.",
author = "Jesper Bonde and Ejegod, {Ditte M{\o}ller} and Kate Cuschieri and Joakim Dillner and Heideman, {Dani{\"e}lle A M} and Wim Quint and {Pavon Ribas}, {Miguel Angel} and Elizaveta Padalko and Christiansen, {Irene Kraus} and Lan Xu and Marc Arbyn",
note = "Copyright {\circledC} 2018 The Authors. Published by Elsevier B.V. All rights reserved.",
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Bonde, J, Ejegod, DM, Cuschieri, K, Dillner, J, Heideman, DAM, Quint, W, Pavon Ribas, MA, Padalko, E, Christiansen, IK, Xu, L & Arbyn, M 2018, 'The Valgent4 protocol: Robust analytical and clinical validation of 11 HPV assays with genotyping on cervical samples collected in SurePath medium' Journal of Clinical Virology, vol. 108, pp. 64-71. https://doi.org/10.1016/j.jcv.2018.09.012

The Valgent4 protocol : Robust analytical and clinical validation of 11 HPV assays with genotyping on cervical samples collected in SurePath medium. / Bonde, Jesper; Ejegod, Ditte Møller; Cuschieri, Kate; Dillner, Joakim; Heideman, Daniëlle A M; Quint, Wim; Pavon Ribas, Miguel Angel; Padalko, Elizaveta; Christiansen, Irene Kraus; Xu, Lan; Arbyn, Marc.

In: Journal of Clinical Virology, Vol. 108, 01.11.2018, p. 64-71.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - The Valgent4 protocol

T2 - Robust analytical and clinical validation of 11 HPV assays with genotyping on cervical samples collected in SurePath medium

AU - Bonde, Jesper

AU - Ejegod, Ditte Møller

AU - Cuschieri, Kate

AU - Dillner, Joakim

AU - Heideman, Daniëlle A M

AU - Quint, Wim

AU - Pavon Ribas, Miguel Angel

AU - Padalko, Elizaveta

AU - Christiansen, Irene Kraus

AU - Xu, Lan

AU - Arbyn, Marc

N1 - Copyright © 2018 The Authors. Published by Elsevier B.V. All rights reserved.

PY - 2018/11/1

Y1 - 2018/11/1

N2 - 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.

AB - 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.

U2 - 10.1016/j.jcv.2018.09.012

DO - 10.1016/j.jcv.2018.09.012

M3 - Article

VL - 108

SP - 64

EP - 71

JO - Journal of Clinical Virology

JF - Journal of Clinical Virology

SN - 1386-6532

ER -