Robust hybridization-based genotyping probes for HPV 6, 11, 16 and 18 obtained via in vitro selection

  • Ivan B. Brukner | Jewish General Hospital, Canada.
  • Anne-Marie Larose
  • Izabella Gorska-Flipot
  • Maja Krajinovic
  • Damian Labuda


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.



PlumX Metrics


Download data is not yet available.

Author Biography

Ivan B. Brukner, Jewish General Hospital
Department of Medical Diagnostics
SMBD-Jewish General Hospital, Suite/Room D-133 & D0-132
3755 Cote-Sainte-Catherine Road, H3T 1E2
Montreal, Quebec, Canada
Tel: 514-3408222 ext 4801 & ext 3699
assay, hybridization, cervical cancer, HPV typing, multiplex assay
  • Abstract views: 1114

  • PDF: 478
  • HTML: 4435
How to Cite
Brukner, I., Larose, A.-M., Gorska-Flipot, I., Krajinovic, M., & Labuda, D. (2010). Robust hybridization-based genotyping probes for HPV 6, 11, 16 and 18 obtained via in vitro selection. Journal of Nucleic Acids Investigation, 1(1), e3.