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Title
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A real-time PCR approach based on SPF10 primers and the INNO-LiPA HPV genotyping extra assay for the detection and typing of human papillomavirus
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Author
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Abstract
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| The SPF10 PCR targets a conserved 65 bp region of the HPV L1 gene for broad-spectrum amplification. The LiPA assay allows subsequent genotyping of the HPV amplicons. This study aims to develop a SPF10 real-time PCR to achieve simultaneous amplification and detection of the HPV target. That way, LiPA analysis of the HPV-negative samples can be avoided, reducing workload and cost. The real-time PCR shows an analytical sensitivity of 29.7 copies for HPV 6, 16, 18 and 31 and an HPV-specific melting peak. Thirty-one HPV DNA plasmids were genotyped correctly using the SPF10 real-time PCR in combination with the LiPA. Here, the LiPA assay was performed at an increased hybridisation temperature (49.5 degrees C) in combination with a reduced amplicon volume (1 mu l) to avoid cross-reactivity. In conclusion, the SPF10 real-time PCR proves to be very sensitive and generates amplicons, which are compatible with the LiPA. (C) 2012 Elsevier B.V. All rights reserved. |
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Language
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English
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Source (journal)
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Journal of virological methods. - Amsterdam
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Publication
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Amsterdam
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2013
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ISSN
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0166-0934
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DOI
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10.1016/J.JVIROMET.2012.09.013
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Volume/pages
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187
:1
(2013)
, p. 166-171
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ISI
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000312609600027
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Full text (Publisher's DOI)
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Full text (publisher's version - intranet only)
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