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 infection.
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.
Snijders
P J
PJ
Department of Pathology, Free University Hospital, Amsterdam, The Netherlands.
Meijer
C J
CJ
Walboomers
J M
JM
D000595:Amino Acid Sequence; D001483:Base Sequence; D015139:Blotting, Southern; D003001:Cloning, Molecular; D004279:DNA, Viral; D006801:Humans; D008969:Molecular Sequence Data; D027383:Papillomaviridae / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013698:Templates, Genetic; D014412:Tumor Virus Infections / Q000175:diagnosis*
D004279:DNA, Viral