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Development and validation of a method for human papillomavirus genotyping based on molecular beacon probes

We describe a new assaying system for the detection and genotyping of human papillomavirus (HPV) based on linear-after-the-exponential-PCR(LATE-PCR) and melting curve analysis. The 23 most prevalent HPV strains (types 6, 11, 16, 18, 31, 33, 35, 39, 42, 45, 51, 52, 53, 56, 58, 59, 66, 68, 70, 73, 81,...

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Autores principales: Lyu, Jiangfeng, Yu, Yuefeng, Pan, Caixia, Zhou, Jing, Ren, Xuyi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6264813/
https://www.ncbi.nlm.nih.gov/pubmed/30496224
http://dx.doi.org/10.1371/journal.pone.0207930
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author Lyu, Jiangfeng
Yu, Yuefeng
Pan, Caixia
Zhou, Jing
Ren, Xuyi
author_facet Lyu, Jiangfeng
Yu, Yuefeng
Pan, Caixia
Zhou, Jing
Ren, Xuyi
author_sort Lyu, Jiangfeng
collection PubMed
description We describe a new assaying system for the detection and genotyping of human papillomavirus (HPV) based on linear-after-the-exponential-PCR(LATE-PCR) and melting curve analysis. The 23 most prevalent HPV strains (types 6, 11, 16, 18, 31, 33, 35, 39, 42, 45, 51, 52, 53, 56, 58, 59, 66, 68, 70, 73, 81, 82, and 83) are assayed in two sealed reaction tubes within 2 h. Good sensitivity and specificity was evaluated by testing cloned HPV DNA and clinical samples. The detection limit was 5–500 copies/reaction depending on the genotype. No cross-reactivity was observed with the other HPV types that are not covered by our method or pathogens tested which were commonly found in female genital tract. When compared with the HPV GenoArray Diagnostic kit, the results from 1104 clinical samples suggest good overall agreement between the two methods,(98.37%, 95% CI: 97.44%–98.97%) and the kappa value was 0.954. Overall, this new HPV genotyping assay system presents a simple, rapid, universally applicable, sensitive, and highly specific detection methodology that should be useful for HPV detection and genotyping, therefore, is potentially of great value in clinical application.
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spelling pubmed-62648132018-12-19 Development and validation of a method for human papillomavirus genotyping based on molecular beacon probes Lyu, Jiangfeng Yu, Yuefeng Pan, Caixia Zhou, Jing Ren, Xuyi PLoS One Research Article We describe a new assaying system for the detection and genotyping of human papillomavirus (HPV) based on linear-after-the-exponential-PCR(LATE-PCR) and melting curve analysis. The 23 most prevalent HPV strains (types 6, 11, 16, 18, 31, 33, 35, 39, 42, 45, 51, 52, 53, 56, 58, 59, 66, 68, 70, 73, 81, 82, and 83) are assayed in two sealed reaction tubes within 2 h. Good sensitivity and specificity was evaluated by testing cloned HPV DNA and clinical samples. The detection limit was 5–500 copies/reaction depending on the genotype. No cross-reactivity was observed with the other HPV types that are not covered by our method or pathogens tested which were commonly found in female genital tract. When compared with the HPV GenoArray Diagnostic kit, the results from 1104 clinical samples suggest good overall agreement between the two methods,(98.37%, 95% CI: 97.44%–98.97%) and the kappa value was 0.954. Overall, this new HPV genotyping assay system presents a simple, rapid, universally applicable, sensitive, and highly specific detection methodology that should be useful for HPV detection and genotyping, therefore, is potentially of great value in clinical application. Public Library of Science 2018-11-29 /pmc/articles/PMC6264813/ /pubmed/30496224 http://dx.doi.org/10.1371/journal.pone.0207930 Text en © 2018 Lyu 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
Lyu, Jiangfeng
Yu, Yuefeng
Pan, Caixia
Zhou, Jing
Ren, Xuyi
Development and validation of a method for human papillomavirus genotyping based on molecular beacon probes
title Development and validation of a method for human papillomavirus genotyping based on molecular beacon probes
title_full Development and validation of a method for human papillomavirus genotyping based on molecular beacon probes
title_fullStr Development and validation of a method for human papillomavirus genotyping based on molecular beacon probes
title_full_unstemmed Development and validation of a method for human papillomavirus genotyping based on molecular beacon probes
title_short Development and validation of a method for human papillomavirus genotyping based on molecular beacon probes
title_sort development and validation of a method for human papillomavirus genotyping based on molecular beacon probes
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6264813/
https://www.ncbi.nlm.nih.gov/pubmed/30496224
http://dx.doi.org/10.1371/journal.pone.0207930
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