Cargando…

Development and interlaboratory agreement of real-time PCR for HPV16 quantification in liquid-based cervical samples

High risk HPV infection is the necessary cause for the development of precancerous and cancerous lesions of the cervix. Among HPV, HPV16 represents the most carcinogenic type. Since the determination of HPV16 DNA load could be clinically useful, we assessed quantitative real-time PCR targeting E6HPV...

Descripción completa

Detalles Bibliográficos
Autores principales: Guenat, David, Dalstein, Véronique, Mauny, Frédéric, Saunier, Maëlle, Briolat, Jenny, Clavel, Christine, Riethmuller, Didier, Mougin, Christiane, Prétet, Jean-Luc
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6202657/
https://www.ncbi.nlm.nih.gov/pubmed/30343011
http://dx.doi.org/10.1016/j.pvr.2018.10.003
_version_ 1783365725196386304
author Guenat, David
Dalstein, Véronique
Mauny, Frédéric
Saunier, Maëlle
Briolat, Jenny
Clavel, Christine
Riethmuller, Didier
Mougin, Christiane
Prétet, Jean-Luc
author_facet Guenat, David
Dalstein, Véronique
Mauny, Frédéric
Saunier, Maëlle
Briolat, Jenny
Clavel, Christine
Riethmuller, Didier
Mougin, Christiane
Prétet, Jean-Luc
author_sort Guenat, David
collection PubMed
description High risk HPV infection is the necessary cause for the development of precancerous and cancerous lesions of the cervix. Among HPV, HPV16 represents the most carcinogenic type. Since the determination of HPV16 DNA load could be clinically useful, we assessed quantitative real-time PCR targeting E6HPV16 and albumin genes on two different platforms. Series of SiHa cells diluted in PreservCyt were used to assess repeatability and reproducibility of two in-house real-time PCR techniques run in two different laboratories to determine HPV16 load. Furthermore, 97 HPV16 positive cervical samples were evaluated to estimate inter-center variability using Bland-Alman plots. As a whole, both techniques presented coefficients of variation for HPV16 load measurement similar to those established for other virus quantification with commercial kits. Moreover, the two real-time PCR techniques showed a very good agreement for HPV16 load calculation. Finally, we emphasize that robust HPV16 DNA quantification requires normalization of viral load by the cell number.
format Online
Article
Text
id pubmed-6202657
institution National Center for Biotechnology Information
language English
publishDate 2018
publisher Elsevier
record_format MEDLINE/PubMed
spelling pubmed-62026572018-10-30 Development and interlaboratory agreement of real-time PCR for HPV16 quantification in liquid-based cervical samples Guenat, David Dalstein, Véronique Mauny, Frédéric Saunier, Maëlle Briolat, Jenny Clavel, Christine Riethmuller, Didier Mougin, Christiane Prétet, Jean-Luc Papillomavirus Res Article High risk HPV infection is the necessary cause for the development of precancerous and cancerous lesions of the cervix. Among HPV, HPV16 represents the most carcinogenic type. Since the determination of HPV16 DNA load could be clinically useful, we assessed quantitative real-time PCR targeting E6HPV16 and albumin genes on two different platforms. Series of SiHa cells diluted in PreservCyt were used to assess repeatability and reproducibility of two in-house real-time PCR techniques run in two different laboratories to determine HPV16 load. Furthermore, 97 HPV16 positive cervical samples were evaluated to estimate inter-center variability using Bland-Alman plots. As a whole, both techniques presented coefficients of variation for HPV16 load measurement similar to those established for other virus quantification with commercial kits. Moreover, the two real-time PCR techniques showed a very good agreement for HPV16 load calculation. Finally, we emphasize that robust HPV16 DNA quantification requires normalization of viral load by the cell number. Elsevier 2018-10-18 /pmc/articles/PMC6202657/ /pubmed/30343011 http://dx.doi.org/10.1016/j.pvr.2018.10.003 Text en © 2018 The Authors http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Article
Guenat, David
Dalstein, Véronique
Mauny, Frédéric
Saunier, Maëlle
Briolat, Jenny
Clavel, Christine
Riethmuller, Didier
Mougin, Christiane
Prétet, Jean-Luc
Development and interlaboratory agreement of real-time PCR for HPV16 quantification in liquid-based cervical samples
title Development and interlaboratory agreement of real-time PCR for HPV16 quantification in liquid-based cervical samples
title_full Development and interlaboratory agreement of real-time PCR for HPV16 quantification in liquid-based cervical samples
title_fullStr Development and interlaboratory agreement of real-time PCR for HPV16 quantification in liquid-based cervical samples
title_full_unstemmed Development and interlaboratory agreement of real-time PCR for HPV16 quantification in liquid-based cervical samples
title_short Development and interlaboratory agreement of real-time PCR for HPV16 quantification in liquid-based cervical samples
title_sort development and interlaboratory agreement of real-time pcr for hpv16 quantification in liquid-based cervical samples
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6202657/
https://www.ncbi.nlm.nih.gov/pubmed/30343011
http://dx.doi.org/10.1016/j.pvr.2018.10.003
work_keys_str_mv AT guenatdavid developmentandinterlaboratoryagreementofrealtimepcrforhpv16quantificationinliquidbasedcervicalsamples
AT dalsteinveronique developmentandinterlaboratoryagreementofrealtimepcrforhpv16quantificationinliquidbasedcervicalsamples
AT maunyfrederic developmentandinterlaboratoryagreementofrealtimepcrforhpv16quantificationinliquidbasedcervicalsamples
AT sauniermaelle developmentandinterlaboratoryagreementofrealtimepcrforhpv16quantificationinliquidbasedcervicalsamples
AT briolatjenny developmentandinterlaboratoryagreementofrealtimepcrforhpv16quantificationinliquidbasedcervicalsamples
AT clavelchristine developmentandinterlaboratoryagreementofrealtimepcrforhpv16quantificationinliquidbasedcervicalsamples
AT riethmullerdidier developmentandinterlaboratoryagreementofrealtimepcrforhpv16quantificationinliquidbasedcervicalsamples
AT mouginchristiane developmentandinterlaboratoryagreementofrealtimepcrforhpv16quantificationinliquidbasedcervicalsamples
AT pretetjeanluc developmentandinterlaboratoryagreementofrealtimepcrforhpv16quantificationinliquidbasedcervicalsamples