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Journal of Clinical Microbiology, September 2001, p. 3204-3212, Vol. 39, No. 9
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.9.3204-3212.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Novel Method for Detection, Typing, and Quantification of Human Papillomaviruses in Clinical Samples

Keith W. Hart,1,* O. Martin Williams,1 Nicola Thelwell,2 Alison N. Fiander,3 Tom Brown,4 Lesek K. Borysiewicz,1,dagger and Colin M. Gelder1

Section of Infection and Immunity, Department of Medicine, University of Wales College of Medicine, Heath Park, Cardiff CF14 4XX,1 Department of Obstetrics and Gynaecology, Llandough Hospital & University Hospital of Wales, Cardiff CF4 4XX,3 Oswel Research Products, Ltd., University of Southampton, Southampton SO16 7PX,2 and Department of Chemistry, University of Southampton, Highfield, Southampton SO17 1BJ,4 United Kingdom

Received 2 April 2001/Returned for modification 20 May 2001/Accepted 16 July 2001

We report the development of a novel detection and typing methodology for human papillomaviruses (HPV) based on real-time PCR with the self-probing fluorescent primers known as Scorpions. This technique is quick, simple, specific, sensitive, and capable of estimating viral load per cell. We report the results of over 100 typing reactions performed on cell lines, biopsies, and cervical cytobrush samples which, when compared to the current reference HPV detection and typing technique, present a kappa  value of 0.89. We further report preliminary data suggesting a relationship between viral load per cell and grade of cervical disease.

* Corresponding author. Mailing address: Section of Infection and Immunity, Department of Medicine, University of Wales College of Medicine, Heath Park, Cardiff CF14 4XX, United Kingdom. Phone: (029) 20745004. Fax: (029) 20745003. E-mail: HartKW{at}cf.ac.uk.

dagger Present address: Imperial College School of Medicine, London SW7 2AZ, United Kingdom.

Journal of Clinical Microbiology, September 2001, p. 3204-3212, Vol. 39, No. 9
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.9.3204-3212.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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