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Development of a sensitive and specific multiplex PCR method combined with DNA microarray primer extension to detect beta-papillomavirus types.

Gheit, Tarik; Billoud, Gaelle; de Koning, Maurits N. C.; Gemignani, Federica; Forslund, Ola LU ; Sylla, Bakary S.; Vaccarella, Salvatore; Franceschi, Silvia; Landi, Stefano and Quint, Wim G. V., et al. (2007) In Journal of Clinical Microbiology 45(8). p.2537-2544
Abstract
Emerging lines of evidence indicate that the cutaneous human papillomavirus (HPV) types that belong to the genus Betapapillomavirus (beta HPV) are involved in the development of nonmelanoma skin cancer. Unlike the situation for mucosal HPV types, highly sensitive and reliable methods to identify characterized cutaneous HPV types in a single assay are limited. Here, we describe a novel one-shot method for the detection of all characterized beta HPV types, namely, HPV type 5 (HPV5), 8, 9, 12, 14, 15, 17, 19, 20, 21, 22, 23, 24, 25, 36, 37, 38, 47, 49, 75, 76, 80, 92, 93, and 96. This assay combines two different techniques: multiplex PCR using HPV type-specific primers for amplification of each E7 gene and array primer extension (APEX) for... (More)
Emerging lines of evidence indicate that the cutaneous human papillomavirus (HPV) types that belong to the genus Betapapillomavirus (beta HPV) are involved in the development of nonmelanoma skin cancer. Unlike the situation for mucosal HPV types, highly sensitive and reliable methods to identify characterized cutaneous HPV types in a single assay are limited. Here, we describe a novel one-shot method for the detection of all characterized beta HPV types, namely, HPV type 5 (HPV5), 8, 9, 12, 14, 15, 17, 19, 20, 21, 22, 23, 24, 25, 36, 37, 38, 47, 49, 75, 76, 80, 92, 93, and 96. This assay combines two different techniques: multiplex PCR using HPV type-specific primers for amplification of each E7 gene and array primer extension (APEX) for typing. This method has been validated using clinical samples which were analyzed simultaneously for the presence of cutaneous HPV types by two additional methods, i.e., the FAP59/64 PCR protocol and a commercially available PCR-reverse hybridization assay (PM-PCR RHA). Our data show good agreement between the results obtained with the multiplex PCR/APEX assay and the PM-PCR RHA method (overall HPV positivity of 92.2% for multiplex PCR/APEX assay versus 90.6% with the PM-PCR RHA) (kappa value, 50; 95% confidence interval, 13 to 88). In addition, the multiplex PCR/APEX assay showed higher sensitivity than the PM-PCR RHA did. This favorable feature and the high-throughput potential make this assay ideal for large-scale clinical and epidemiological studies aimed at determining the spectrum of cutaneous types in skin cancer. (Less)
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organization
publishing date
type
Contribution to journal
publication status
published
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in
Journal of Clinical Microbiology
volume
45
issue
8
pages
2537 - 2544
publisher
American Society for Microbiology
external identifiers
  • wos:000248793300028
  • scopus:34548061824
ISSN
1098-660X
DOI
10.1128/JCM.00747-07
language
English
LU publication?
yes
id
47c14925-40f5-4860-99fc-8065190c8002 (old id 691771)
date added to LUP
2007-12-11 21:14:17
date last changed
2017-10-29 04:16:20
@article{47c14925-40f5-4860-99fc-8065190c8002,
  abstract     = {Emerging lines of evidence indicate that the cutaneous human papillomavirus (HPV) types that belong to the genus Betapapillomavirus (beta HPV) are involved in the development of nonmelanoma skin cancer. Unlike the situation for mucosal HPV types, highly sensitive and reliable methods to identify characterized cutaneous HPV types in a single assay are limited. Here, we describe a novel one-shot method for the detection of all characterized beta HPV types, namely, HPV type 5 (HPV5), 8, 9, 12, 14, 15, 17, 19, 20, 21, 22, 23, 24, 25, 36, 37, 38, 47, 49, 75, 76, 80, 92, 93, and 96. This assay combines two different techniques: multiplex PCR using HPV type-specific primers for amplification of each E7 gene and array primer extension (APEX) for typing. This method has been validated using clinical samples which were analyzed simultaneously for the presence of cutaneous HPV types by two additional methods, i.e., the FAP59/64 PCR protocol and a commercially available PCR-reverse hybridization assay (PM-PCR RHA). Our data show good agreement between the results obtained with the multiplex PCR/APEX assay and the PM-PCR RHA method (overall HPV positivity of 92.2% for multiplex PCR/APEX assay versus 90.6% with the PM-PCR RHA) (kappa value, 50; 95% confidence interval, 13 to 88). In addition, the multiplex PCR/APEX assay showed higher sensitivity than the PM-PCR RHA did. This favorable feature and the high-throughput potential make this assay ideal for large-scale clinical and epidemiological studies aimed at determining the spectrum of cutaneous types in skin cancer.},
  author       = {Gheit, Tarik and Billoud, Gaelle and de Koning, Maurits N. C. and Gemignani, Federica and Forslund, Ola and Sylla, Bakary S. and Vaccarella, Salvatore and Franceschi, Silvia and Landi, Stefano and Quint, Wim G. V. and Canzian, Federico and Tommasino, Massimo},
  issn         = {1098-660X},
  language     = {eng},
  number       = {8},
  pages        = {2537--2544},
  publisher    = {American Society for Microbiology},
  series       = {Journal of Clinical Microbiology},
  title        = {Development of a sensitive and specific multiplex PCR method combined with DNA microarray primer extension to detect beta-papillomavirus types.},
  url          = {http://dx.doi.org/10.1128/JCM.00747-07},
  volume       = {45},
  year         = {2007},
}