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(American Journal of Pathology. 1998;153:1731-1739.)
© 1998 American Society for Investigative Pathology

Technical Advance

Novel Short-Fragment PCR Assay for Highly Sensitive Broad-Spectrum Detection of Anogenital Human Papillomaviruses

Bernhard Kleter* , Leen-Jan van Doorn* , Jan ter Schegget , Lianne Schrauwen* , Kees van Krimpen , Matthé Burger§ , Bram ter Harmsel¶ and Wim Quint*

From the Delft Diagnostic Laboratory,* the Department of Pathology, Diagnostic Center SSDZ, and the Department of Gynecology,¶ and the Departments of Virology and Gynecology,§ Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands

A novel set of polymerase chain reaction (PCR) primers, designated SPF1 and SPF2 and located in the L1 region, was developed for universal detection of human papillomavirus (HPV). A short PCR fragment (SPF) of only 65 pb was synthesized. SPF amplimers were detected in a microtiter-based hybridization system, using a mixture of oligonucleotide probes. The SPF system allowed detection of at least 43 different HPV genotypes. The clinical performance of the novel SPF system was assessed in three different patient groups. 1) Analysis of 534 cervical scrapes, obtained from treated patients, showed that the detection rate in 447 (83.7%) scrapes with normal cytology was significantly higher using the SPF system as compared with the universal primer set GP5+/6+ (P < 0.001). 2) The SPF assay detected HPV DNA in 299 (98.4%) of 304 scrapes with cytological dyskaryosis. 3) The SPF system detected HPV DNA in 100% of 184 formalin-fixed, paraffin-embedded cervical carcinoma specimens. In conclusion, the novel SPF system permitted universal and highly sensitive detection of HPV DNA in diverse clinical materials and may improve the molecular diagnosis and epidemiology of this important virus.

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