Cargando…

Methylation analysis and HPV genotyping of self-collected cervical samples from women not responding to screening invitation and review of the literature

AIM OF THE STUDY: To assess the feasibility of partial HPV genotyping and methylation analysis of CADM1, MAL, and miR124-2 genes as triage tests in assaying self-collected cervical samples positive for high-risk HPV on primary screening, and to review the literature regarding host cellular gene meth...

Descripción completa

Detalles Bibliográficos
Autores principales: Del Mistro, Annarosa, Frayle, Helena, Rizzi, Martina, Fantin, Gianpiero, Ferro, Antonio, Angeletti, Paolo Matteo, Giorgi Rossi, Paolo, Altobelli, Emma
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5338782/
https://www.ncbi.nlm.nih.gov/pubmed/28263992
http://dx.doi.org/10.1371/journal.pone.0172226
_version_ 1782512552600666112
author Del Mistro, Annarosa
Frayle, Helena
Rizzi, Martina
Fantin, Gianpiero
Ferro, Antonio
Angeletti, Paolo Matteo
Giorgi Rossi, Paolo
Altobelli, Emma
author_facet Del Mistro, Annarosa
Frayle, Helena
Rizzi, Martina
Fantin, Gianpiero
Ferro, Antonio
Angeletti, Paolo Matteo
Giorgi Rossi, Paolo
Altobelli, Emma
author_sort Del Mistro, Annarosa
collection PubMed
description AIM OF THE STUDY: To assess the feasibility of partial HPV genotyping and methylation analysis of CADM1, MAL, and miR124-2 genes as triage tests in assaying self-collected cervical samples positive for high-risk HPV on primary screening, and to review the literature regarding host cellular gene methylation analysis of self-collected cervical samples. MATERIAL AND METHODS: Women residing in North-East Italy who had failed to respond to the invitation to participate in an organized population-based program were invited to provide a self-sample. Their stored baseline (self-collected) and follow-up (clinician-collected) cervical samples were included in the study. DNA was extracted from HPV-positive (Qiagen’s Hybrid Capture 2, HC2) samples. Partial genotyping with separate detection of HPV types 16 and 18 was performed with a hybrid capture-based method and a quantitative PCR assay. Methylation was assayed with a quantitative methylation-specific PCR. RESULTS: High-risk HPV infection was detected in 48% of baseline and 71% of follow-up HC2-positive samples. Methylation was demonstrated respectively in 15% and 23.5% of baseline and follow-up samples and chiefly involved a single gene (miR124-2). Invalid quantitative PCR results were recorded in 5% of self-collected samples. The specificity of miR124-1, MAL, and CADM1 methylation was 84%, 94%, and 98%, respectively, and the specificity of the three markers combined was 84%. Sensitivity was not estimated due to the lack of CIN2+ samples. The systematic review showed that different methylation assays yield different accuracy values. CONCLUSION: Self-collected samples are suitable for methylation assays included in reflex triage testing. The reproducibility and accuracy of the methylation tests described in the literature should be improved.
format Online
Article
Text
id pubmed-5338782
institution National Center for Biotechnology Information
language English
publishDate 2017
publisher Public Library of Science
record_format MEDLINE/PubMed
spelling pubmed-53387822017-03-10 Methylation analysis and HPV genotyping of self-collected cervical samples from women not responding to screening invitation and review of the literature Del Mistro, Annarosa Frayle, Helena Rizzi, Martina Fantin, Gianpiero Ferro, Antonio Angeletti, Paolo Matteo Giorgi Rossi, Paolo Altobelli, Emma PLoS One Research Article AIM OF THE STUDY: To assess the feasibility of partial HPV genotyping and methylation analysis of CADM1, MAL, and miR124-2 genes as triage tests in assaying self-collected cervical samples positive for high-risk HPV on primary screening, and to review the literature regarding host cellular gene methylation analysis of self-collected cervical samples. MATERIAL AND METHODS: Women residing in North-East Italy who had failed to respond to the invitation to participate in an organized population-based program were invited to provide a self-sample. Their stored baseline (self-collected) and follow-up (clinician-collected) cervical samples were included in the study. DNA was extracted from HPV-positive (Qiagen’s Hybrid Capture 2, HC2) samples. Partial genotyping with separate detection of HPV types 16 and 18 was performed with a hybrid capture-based method and a quantitative PCR assay. Methylation was assayed with a quantitative methylation-specific PCR. RESULTS: High-risk HPV infection was detected in 48% of baseline and 71% of follow-up HC2-positive samples. Methylation was demonstrated respectively in 15% and 23.5% of baseline and follow-up samples and chiefly involved a single gene (miR124-2). Invalid quantitative PCR results were recorded in 5% of self-collected samples. The specificity of miR124-1, MAL, and CADM1 methylation was 84%, 94%, and 98%, respectively, and the specificity of the three markers combined was 84%. Sensitivity was not estimated due to the lack of CIN2+ samples. The systematic review showed that different methylation assays yield different accuracy values. CONCLUSION: Self-collected samples are suitable for methylation assays included in reflex triage testing. The reproducibility and accuracy of the methylation tests described in the literature should be improved. Public Library of Science 2017-03-06 /pmc/articles/PMC5338782/ /pubmed/28263992 http://dx.doi.org/10.1371/journal.pone.0172226 Text en © 2017 Del Mistro et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Del Mistro, Annarosa
Frayle, Helena
Rizzi, Martina
Fantin, Gianpiero
Ferro, Antonio
Angeletti, Paolo Matteo
Giorgi Rossi, Paolo
Altobelli, Emma
Methylation analysis and HPV genotyping of self-collected cervical samples from women not responding to screening invitation and review of the literature
title Methylation analysis and HPV genotyping of self-collected cervical samples from women not responding to screening invitation and review of the literature
title_full Methylation analysis and HPV genotyping of self-collected cervical samples from women not responding to screening invitation and review of the literature
title_fullStr Methylation analysis and HPV genotyping of self-collected cervical samples from women not responding to screening invitation and review of the literature
title_full_unstemmed Methylation analysis and HPV genotyping of self-collected cervical samples from women not responding to screening invitation and review of the literature
title_short Methylation analysis and HPV genotyping of self-collected cervical samples from women not responding to screening invitation and review of the literature
title_sort methylation analysis and hpv genotyping of self-collected cervical samples from women not responding to screening invitation and review of the literature
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5338782/
https://www.ncbi.nlm.nih.gov/pubmed/28263992
http://dx.doi.org/10.1371/journal.pone.0172226
work_keys_str_mv AT delmistroannarosa methylationanalysisandhpvgenotypingofselfcollectedcervicalsamplesfromwomennotrespondingtoscreeninginvitationandreviewoftheliterature
AT fraylehelena methylationanalysisandhpvgenotypingofselfcollectedcervicalsamplesfromwomennotrespondingtoscreeninginvitationandreviewoftheliterature
AT rizzimartina methylationanalysisandhpvgenotypingofselfcollectedcervicalsamplesfromwomennotrespondingtoscreeninginvitationandreviewoftheliterature
AT fantingianpiero methylationanalysisandhpvgenotypingofselfcollectedcervicalsamplesfromwomennotrespondingtoscreeninginvitationandreviewoftheliterature
AT ferroantonio methylationanalysisandhpvgenotypingofselfcollectedcervicalsamplesfromwomennotrespondingtoscreeninginvitationandreviewoftheliterature
AT angelettipaolomatteo methylationanalysisandhpvgenotypingofselfcollectedcervicalsamplesfromwomennotrespondingtoscreeninginvitationandreviewoftheliterature
AT giorgirossipaolo methylationanalysisandhpvgenotypingofselfcollectedcervicalsamplesfromwomennotrespondingtoscreeninginvitationandreviewoftheliterature
AT altobelliemma methylationanalysisandhpvgenotypingofselfcollectedcervicalsamplesfromwomennotrespondingtoscreeninginvitationandreviewoftheliterature