%0 Journal Article
%T Robust hybridization-based genotyping probes for HPV 6, 11, 16 and 18 obtained via in vitro selection
%A Ivan B. Brukner
%A Anne-Marie Larose
%A Izabella Gorska-Flipot
%A Maja Krajinovic
%J Journal of Nucleic Acids Investigation
%D 2010
%I PAGEPress Publications
%R 10.4081/jnai.2010.e3
%X This paper describes the technical and analytical performance of a novel set of hybridization probes for the four GARDASIL  vaccine-relevant HPV types (6, 11, 16 and 18). These probes are obtained through in vitro selection from a pool of random oligonucleotides, rather than the traditional ¡°rational design¡± approach typically used as the initial step in assay development. The type-specific segment of the HPV genome was amplified using a GP5+/6+ PCR protocol and 39 synthetic oligonucleotide templates derived from each of the HPV types, as PCR targets. The robust performance of the 4 selected hybridization probes was demonstrated by monitoring the preservation of the specificity and sensitivity of the typing assay over all 39 HPV types, using a different spectrum of HPV (genome equivalent: 103-109) and human DNA concentrations (10-100 ng) as well as temperature and buffer composition variations. To the Authors¡¯ knowledge, this is a unique hybridization-based multiplex typing assay. It performs at ambient temperatures, does not require the strict temperature control of hybridization conditions, and is functional with a number of different non-denaturing buffers, thereby offering downstream compatibility with a variety of detection methods. Studies aimed at demonstrating clinical performance are needed to validate the applicability of this strategy.
%K assay
%K hybridization
%K cervical cancer
%K HPV typing
%K multiplex assay
%U http://www.pagepress.org/journals/index.php/jnai/article/view/1491