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Degenerate primers based on highly conserved regions of amino acid sequence in papillomaviruses can be used in a generalized polymerase chain reaction to detect productive human papillomavirus infections

Department of Pathology, Section of Molecular Pathology, Free University Hospital, De Boelelaan 1117, 1081 HV Amsterdam, The Netherlands

Conserved amino acid sequences within the L1 open reading frame of the human papillomavirus (HPV) genome were used as a basis to design two degenerate primers (GP17 and GP18) and one general probe (GPR22) which direct polymerase chain reaction (PCR) amplification and subsequent detection of a 620 to 660 bp DNA fragment. The conserved nature of the primers and probe was tested experimentally on a panel of 24 cloned HPV DNAs isolated from cutaneous and mucosal lesions, including HPV-2a and -57, which are known to be associated with lesions at both anatomical sites. The sensitivity of this PCR test was at the level of genomic Southern blot analysis, indicating that HPV infections producing high copy numbers can be detected. Positive results were obtained with DNA extracted from clinical samples of genital and cutaneous origin.

Received 8 April 1991; accepted 17 July 1991.

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