High-resolution melting molecular signatures for rapid identification of human papillomavirus genotypes.

BACKGROUND: Genotyping of human papillomarvirus (HPV) is crucial for patient management in a clinical setting. This study accesses the combined use of broad-range real-time PCR and high-resolution melting (HRM) analysis for rapid identification of HPV genotypes. METHODS: Genomic DNA sequences of 8 h...

Full description

Bibliographic Details
Main Authors: Ta-Hsien Lee, Tzong-Shoon Wu, Ching-Ping Tseng, Jiantai Timothy Qiu
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2012-01-01
Series:PLoS ONE
Online Access:http://europepmc.org/articles/PMC3423390?pdf=render
_version_ 1818498046716018688
author Ta-Hsien Lee
Tzong-Shoon Wu
Ching-Ping Tseng
Jiantai Timothy Qiu
author_facet Ta-Hsien Lee
Tzong-Shoon Wu
Ching-Ping Tseng
Jiantai Timothy Qiu
author_sort Ta-Hsien Lee
collection DOAJ
description BACKGROUND: Genotyping of human papillomarvirus (HPV) is crucial for patient management in a clinical setting. This study accesses the combined use of broad-range real-time PCR and high-resolution melting (HRM) analysis for rapid identification of HPV genotypes. METHODS: Genomic DNA sequences of 8 high-risk genotypes (HPV16/18/39/45/52/56/58/68) were subject to bioinformatic analysis to select for appropriate PCR amplicon. Asymmetric broad-range real-time PCR in the presence of HRM dye and two unlabeled probes specific to HPV16 and 18 was employed to generate HRM molecular signatures for HPV genotyping. The method was validated via assessment of 119 clinical HPV isolates. RESULTS: A DNA fragment within the L1 region was selected as the PCR amplicon ranging from 215-221 bp for different HPV genotypes. Each genotype displayed a distinct HRM molecular signature with minimal inter-assay variability. According to the HRM molecular signatures, HPV genotypes can be determined with one PCR within 3 h from the time of viral DNA isolation. In the validation assay, a 91% accuracy rate was achieved when the genotypes were in the database. Concomitantly, the HRM molecular signatures for additional 6 low-risk genotypes were established. CONCLUSIONS: This assay provides a novel approach for HPV genotyping in a rapid and cost-effective manner.
first_indexed 2024-12-10T18:52:58Z
format Article
id doaj.art-d1dd751b033d42bdbc5fb4b9a925c8cb
institution Directory Open Access Journal
issn 1932-6203
language English
last_indexed 2024-12-10T18:52:58Z
publishDate 2012-01-01
publisher Public Library of Science (PLoS)
record_format Article
series PLoS ONE
spelling doaj.art-d1dd751b033d42bdbc5fb4b9a925c8cb2022-12-22T01:37:16ZengPublic Library of Science (PLoS)PLoS ONE1932-62032012-01-0178e4205110.1371/journal.pone.0042051High-resolution melting molecular signatures for rapid identification of human papillomavirus genotypes.Ta-Hsien LeeTzong-Shoon WuChing-Ping TsengJiantai Timothy QiuBACKGROUND: Genotyping of human papillomarvirus (HPV) is crucial for patient management in a clinical setting. This study accesses the combined use of broad-range real-time PCR and high-resolution melting (HRM) analysis for rapid identification of HPV genotypes. METHODS: Genomic DNA sequences of 8 high-risk genotypes (HPV16/18/39/45/52/56/58/68) were subject to bioinformatic analysis to select for appropriate PCR amplicon. Asymmetric broad-range real-time PCR in the presence of HRM dye and two unlabeled probes specific to HPV16 and 18 was employed to generate HRM molecular signatures for HPV genotyping. The method was validated via assessment of 119 clinical HPV isolates. RESULTS: A DNA fragment within the L1 region was selected as the PCR amplicon ranging from 215-221 bp for different HPV genotypes. Each genotype displayed a distinct HRM molecular signature with minimal inter-assay variability. According to the HRM molecular signatures, HPV genotypes can be determined with one PCR within 3 h from the time of viral DNA isolation. In the validation assay, a 91% accuracy rate was achieved when the genotypes were in the database. Concomitantly, the HRM molecular signatures for additional 6 low-risk genotypes were established. CONCLUSIONS: This assay provides a novel approach for HPV genotyping in a rapid and cost-effective manner.http://europepmc.org/articles/PMC3423390?pdf=render
spellingShingle Ta-Hsien Lee
Tzong-Shoon Wu
Ching-Ping Tseng
Jiantai Timothy Qiu
High-resolution melting molecular signatures for rapid identification of human papillomavirus genotypes.
PLoS ONE
title High-resolution melting molecular signatures for rapid identification of human papillomavirus genotypes.
title_full High-resolution melting molecular signatures for rapid identification of human papillomavirus genotypes.
title_fullStr High-resolution melting molecular signatures for rapid identification of human papillomavirus genotypes.
title_full_unstemmed High-resolution melting molecular signatures for rapid identification of human papillomavirus genotypes.
title_short High-resolution melting molecular signatures for rapid identification of human papillomavirus genotypes.
title_sort high resolution melting molecular signatures for rapid identification of human papillomavirus genotypes
url http://europepmc.org/articles/PMC3423390?pdf=render
work_keys_str_mv AT tahsienlee highresolutionmeltingmolecularsignaturesforrapididentificationofhumanpapillomavirusgenotypes
AT tzongshoonwu highresolutionmeltingmolecularsignaturesforrapididentificationofhumanpapillomavirusgenotypes
AT chingpingtseng highresolutionmeltingmolecularsignaturesforrapididentificationofhumanpapillomavirusgenotypes
AT jiantaitimothyqiu highresolutionmeltingmolecularsignaturesforrapididentificationofhumanpapillomavirusgenotypes