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Journal of Clinical Microbiology, March 2007, p. 928-934, Vol. 45, No. 3
0095-1137/07/$08.00+0     doi:10.1128/JCM.02098-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Evaluation of Combined General Primer-Mediated PCR Sequencing and Type-Specific PCR Strategies for Determination of Human Papillomavirus Genotypes in Cervical Cell Specimens

Véronique Fontaine,^1* Corinne Mascaux,² Christine Weyn,¹ Aurore Bernis,¹ Nathalie Celio,¹ Philippe Lefèvre,¹ Leonard Kaufman,³ and Christian Garbar^2,

Laboratory of Molecular Virology, ISP/Institut Pasteur, rue Engeland 642, 1180 Brussels, Belgium,¹ Department of Pathology, CHU Charleroi, Boulevard Zoé Drion 1, 6000 Charleroi, Belgium,² Department of Biostatistics and Medical Informatics, Brussels Free University/VUB, Brussels 1090, Belgium³

Received 12 October 2006/ Returned for modification 19 December 2006/ Accepted 5 January 2007

A strategy combining human papillomavirus general primer (mainly the PGMY primers)-directed PCR sequencing and type-specific PCR is presented. DNA samples were first tested in general primer-mediated PCR. The amplified fragments of positive samples after ethidium bromide-stained DNA gel analysis were further sequenced, and corresponding DNA samples were further analyzed by PCR using type-specific primers for human papillomavirus (HPV) types 16, 18, 31, and 52. The comparison of the results of 157 samples analyzed by this strategy in parallel with the Hybrid Capture 2 tests and with the HPV INNO-LiPA (Innogenetics line probe assay) shows that this method is suitable for HPV detection and genotyping in cervical cell samples. Although the PCR sequencing method is as sensitive as the HPV INNO-LiPA for HPV detection, our method allows the identification of a broader range of HPV types. In contrast, the HPV INNO-LiPA was less time-consuming and better identified coinfections.

Published ahead of print on 17 January 2007.

Present address: Department of Pathology, AZ-Brussels Free University/VUB, Brussels 1090, Belgium.