PCR-RFLP assay as an option for primary HPV test

PCR-RFLP assay as an option for primary HPV test

Persistent human papillomavirus (HPV) infection is an essential factor of cervical cancer. This study evaluated the analytical performance of restriction fragment length polymorphism polymerase chain reaction (PCR-RFLP) assay compared to PapilloCheck® microarray to identify human papilloma virus (HPV) in cervical cells. Three hundred and twenty-five women were analyzed. One sample was used for conventional cytology and another sample was collected using BD SurePath™ kit for HPV tests. Eighty samples (24.6%) were positive for HPV gene by PCR-Multiplex and were then submitted to PCR-RFLP and PapilloCheck® microarray. There was a genotyping agreement in 71.25% (57/80) on at least one HPV type between PCR-RFLP and PapilloCheck® microarray. In 22 samples (27.5%), the results were discordant and those samples were additionally analyzed by DNA sequencing. HPV 16 was the most prevalent HPV type found in both methods, followed by HPVs 53, 68, 18, 39, and 66 using PCR-RFLP analysis, and HPVs 39, 53, 68, 56, 31, and 66 using PapilloCheck® microarray. In the present study, a perfect agreement using Cohen's kappa (κ) was found in HPV 33 and 58 (κ=1), very good for HPV 51, and good for types 16, 18, 53, 59, 66, 68, 70, and 73. PCR-RFLP analysis identified only 25% (20/80) HPV coinfection, and PapilloCheck® microarray found 62.5% (50/80). Our Cohen's kappa results indicate that our in-house HPV genotyping testing (PCR-RFLP analysis) could be applied as a primary HPV test screening, especially in low income countries. If multiple HPV types are found in this primary test, a more descriptive test, such as PapilloCheck® microarray, could be performed.

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Main Authors: Golfetto,L., Alves,E.V., Martins,T.R., Sincero,T.C.M., Castro,J.B.S., Dannebrock,C., Oliveira,J.G., Levi,J.E., Onofre,A.S.C., Bazzo,M.L.
Format: Digital revista
Language:English
Published: Associação Brasileira de Divulgação Científica 2018
Online Access:http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2018000500619
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spelling oai:scielo:S0100-879X20180005006192019-03-19PCR-RFLP assay as an option for primary HPV testGolfetto,L.Alves,E.V.Martins,T.R.Sincero,T.C.M.Castro,J.B.S.Dannebrock,C.Oliveira,J.G.Levi,J.E.Onofre,A.S.C.Bazzo,M.L. Cervical cancer HPV Screening PCR-RFLP PapilloCheck® microarray Persistent human papillomavirus (HPV) infection is an essential factor of cervical cancer. This study evaluated the analytical performance of restriction fragment length polymorphism polymerase chain reaction (PCR-RFLP) assay compared to PapilloCheck® microarray to identify human papilloma virus (HPV) in cervical cells. Three hundred and twenty-five women were analyzed. One sample was used for conventional cytology and another sample was collected using BD SurePath™ kit for HPV tests. Eighty samples (24.6%) were positive for HPV gene by PCR-Multiplex and were then submitted to PCR-RFLP and PapilloCheck® microarray. There was a genotyping agreement in 71.25% (57/80) on at least one HPV type between PCR-RFLP and PapilloCheck® microarray. In 22 samples (27.5%), the results were discordant and those samples were additionally analyzed by DNA sequencing. HPV 16 was the most prevalent HPV type found in both methods, followed by HPVs 53, 68, 18, 39, and 66 using PCR-RFLP analysis, and HPVs 39, 53, 68, 56, 31, and 66 using PapilloCheck® microarray. In the present study, a perfect agreement using Cohen's kappa (κ) was found in HPV 33 and 58 (κ=1), very good for HPV 51, and good for types 16, 18, 53, 59, 66, 68, 70, and 73. PCR-RFLP analysis identified only 25% (20/80) HPV coinfection, and PapilloCheck® microarray found 62.5% (50/80). Our Cohen's kappa results indicate that our in-house HPV genotyping testing (PCR-RFLP analysis) could be applied as a primary HPV test screening, especially in low income countries. If multiple HPV types are found in this primary test, a more descriptive test, such as PapilloCheck® microarray, could be performed.info:eu-repo/semantics/openAccessAssociação Brasileira de Divulgação CientíficaBrazilian Journal of Medical and Biological Research v.51 n.5 20182018-01-01info:eu-repo/semantics/articletext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2018000500619en10.1590/1414-431x20177098
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language English
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author Golfetto,L.
Alves,E.V.
Martins,T.R.
Sincero,T.C.M.
Castro,J.B.S.
Dannebrock,C.
Oliveira,J.G.
Levi,J.E.
Onofre,A.S.C.
Bazzo,M.L.
spellingShingle Golfetto,L.
Alves,E.V.
Martins,T.R.
Sincero,T.C.M.
Castro,J.B.S.
Dannebrock,C.
Oliveira,J.G.
Levi,J.E.
Onofre,A.S.C.
Bazzo,M.L.
PCR-RFLP assay as an option for primary HPV test
author_facet Golfetto,L.
Alves,E.V.
Martins,T.R.
Sincero,T.C.M.
Castro,J.B.S.
Dannebrock,C.
Oliveira,J.G.
Levi,J.E.
Onofre,A.S.C.
Bazzo,M.L.
author_sort Golfetto,L.
title PCR-RFLP assay as an option for primary HPV test
title_short PCR-RFLP assay as an option for primary HPV test
title_full PCR-RFLP assay as an option for primary HPV test
title_fullStr PCR-RFLP assay as an option for primary HPV test
title_full_unstemmed PCR-RFLP assay as an option for primary HPV test
title_sort pcr-rflp assay as an option for primary hpv test
description Persistent human papillomavirus (HPV) infection is an essential factor of cervical cancer. This study evaluated the analytical performance of restriction fragment length polymorphism polymerase chain reaction (PCR-RFLP) assay compared to PapilloCheck® microarray to identify human papilloma virus (HPV) in cervical cells. Three hundred and twenty-five women were analyzed. One sample was used for conventional cytology and another sample was collected using BD SurePath™ kit for HPV tests. Eighty samples (24.6%) were positive for HPV gene by PCR-Multiplex and were then submitted to PCR-RFLP and PapilloCheck® microarray. There was a genotyping agreement in 71.25% (57/80) on at least one HPV type between PCR-RFLP and PapilloCheck® microarray. In 22 samples (27.5%), the results were discordant and those samples were additionally analyzed by DNA sequencing. HPV 16 was the most prevalent HPV type found in both methods, followed by HPVs 53, 68, 18, 39, and 66 using PCR-RFLP analysis, and HPVs 39, 53, 68, 56, 31, and 66 using PapilloCheck® microarray. In the present study, a perfect agreement using Cohen's kappa (κ) was found in HPV 33 and 58 (κ=1), very good for HPV 51, and good for types 16, 18, 53, 59, 66, 68, 70, and 73. PCR-RFLP analysis identified only 25% (20/80) HPV coinfection, and PapilloCheck® microarray found 62.5% (50/80). Our Cohen's kappa results indicate that our in-house HPV genotyping testing (PCR-RFLP analysis) could be applied as a primary HPV test screening, especially in low income countries. If multiple HPV types are found in this primary test, a more descriptive test, such as PapilloCheck® microarray, could be performed.
publisher Associação Brasileira de Divulgação Científica
publishDate 2018
url http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2018000500619
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