Detection and differentiation of human papillomavirus genotypes HPV-6 and HPV-11 by FRET-based real-time PCR

Journal of Virological Methods
Bostjan J KocjanMario Poljak

Abstract

A real-time PCR (RT-PCR) assay was developed based on fluorescence resonance energy transfer (FRET) hybridization probe technology, allowing very sensitive and specific detection of HPV-6 and HPV-11, reliable differentiation of HPV-6 and HPV-11, as well as prototypic and non-prototypic HPV-6 genomic variants, in a single PCR reaction. The primers and probe were designed on the basis of multiple alignments of 74 HPV-6 E2 gene sequences and 20 HPV-11 E2 gene sequences. Testing on defined plasmid standards showed that the RT-PCR allowed simple and reliable identification of HPV-6 and HPV-11 using type specific amplification followed by probe-specific post-amplification dissociation analysis. Sensitivity, assessed by probit analysis at a 95% detection level, was 42.9, 43.4, and 25.3 DNA copies per assay for prototypic and non-prototypic HPV-6 variants and HPV-11, respectively. The results obtained by the developed assay on 51 HPV DNA-negative samples and 149 HPV DNA-positive samples, including 81 HPV-6 positive samples (19 prototypic and 62 non-prototypic HPV-6 variants), 28 HPV-11 positive samples, 10 samples of HPV-44 and HPV-74 (the closest relatives of HPV-6 and HPV-11) and 30 samples of 15 other important alpha HPV, showed com...Continue Reading

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Citations

Jul 9, 2010·Clinical Microbiology Reviews·Eileen M Burd
Jun 6, 2009·Journal of Clinical Microbiology·Mario PoljakAdriana Vince
Jun 27, 2014·European Archives of Oto-rhino-laryngology : Official Journal of the European Federation of Oto-Rhino-Laryngological Societies (EUFOS) : Affiliated with the German Society for Oto-Rhino-Laryngology - Head and Neck Surgery·Irena Hočevar-BoltežarMiha Zargi
Mar 1, 2009·Expert Opinion on Medical Diagnostics·George KoliopoulosEvgenia Zilakou
Mar 12, 2013·Journal of Virological Methods·Catharina E CombrinckFelicity J Burt
Jan 31, 2012·European Journal of Internal Medicine·Afshin AbdiradNader Sadigh
Oct 31, 2015·Journal of Clinical Virology : the Official Publication of the Pan American Society for Clinical Virology·Boštjan J KocjanMario Poljak
Oct 15, 2013·Clinical Microbiology and Infection : the Official Publication of the European Society of Clinical Microbiology and Infectious Diseases·J M GodínezI G Bravo
Dec 21, 2014·The British Journal of Ophthalmology·Jernej MlakarMario Poljak
Nov 24, 2012·Journal of Virological Methods·Lea HošnjakMario Poljak
May 8, 2017·OTO Open : the Official Open Access Journal of the American Academy of Otolaryngology--Head and Neck Surgery Foundation·Mohammed Abdel Motaal GomaaKhalid M Mohiy

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