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

M Isabel Micalessi; Gaëlle A Boulet; Johannes Bogers

doi:10.1007/978-1-4939-2013-6_2

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

Methods Mol Biol. 2015:1249:27-35. doi: 10.1007/978-1-4939-2013-6_2.

Authors

M Isabel Micalessi¹, Gaëlle A Boulet, Johannes Bogers

Affiliation

¹ Applied Molecular Biology Research (AMBIOR) group, Laboratory of Cell Biology and Histology, Vaccine & Infectious Diseases Institute (VAXINFECTIO), University of Antwerp (Campus Groenenborger), Groenenborgerlaan 171, B-2020, Antwerp, Belgium, [email protected].

PMID: 25348295
DOI: 10.1007/978-1-4939-2013-6_2

Abstract

A highly sensitive SPF10 real-time PCR was developed to achieve simultaneous amplification and detection of the human papillomavirus (HPV) target. That way, LiPA analysis of the HPV-negative samples can be avoided, reducing workload and cost. Here, we describe in detail a SYBR Green I-based real-time PCR assay based on SPF10 primers using the LightCycler(®) 480 system to generate and detect HPV amplicons, which are compatible with the LiPA assay.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

DNA Primers / metabolism*
Genotype
Genotyping Techniques / methods*
Humans
Molecular Typing / methods*
Nucleic Acid Denaturation / genetics
Papillomaviridae / classification*
Papillomaviridae / genetics*
Papillomaviridae / isolation & purification
Real-Time Polymerase Chain Reaction / methods*

Substances

DNA Primers