title: Thermus aquaticus DNA polymerase-catalysed chain reaction for the detection of human papillomaviruses.
abstract: To delineate the conditions for the polymerase chain reaction (PCR) using primers specific for human papillomavirus (HPV) types 6b, 16 and 18, a number of important technical features were analysed. Buffer, concentrations of magnesium, Taq polymerase, primers and DNA templates, annealing temperature, and extension time were studied by a combination of gel electrophoresis, Southern and slot-blot hybridization. Amplification of E6 gene fragments of HPV-16 and HPV-18 generated bands of 110 bp and 154 bp respectively, as predicted. However, amplification of a segment within the long control region of HPV 6b yielded an unexpected size of 340 bp. Different conditions were found for each HPV type-specific primer pair. These results, and the applications of PCR in HPV research and in an increasingly wide range of fields in medical virology are discussed.
authors.affiliation: Institute of Molecular and Cell Biology, National University of Singapore.
authors.forename: V T
authors.initials: VT
authors.lastname: Chow
authors.forename: K M
authors.initials: KM
authors.lastname: Tham
authors.forename: H U
authors.initials: HU
authors.lastname: Bernard
mesh_terms: D001483:Base Sequence; D015139:Blotting, Southern; D002460:Cell Line; D004279:DNA, Viral / Q000096:biosynthesis; D004259:DNA-Directed DNA Polymerase; D006801:Humans; D008969:Molecular Sequence Data; D021141:Nucleic Acid Amplification Techniques; D015345:Oligonucleotide Probes; D027383:Papillomaviridae / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods*; D019914:Taq Polymerase; D013696:Temperature; D013997:Time Factors; D014412:Tumor Virus Infections / Q000175:diagnosis*
chemical_list: D004279:DNA, Viral; D015345:Oligonucleotide Probes; D019914:Taq Polymerase; D004259:DNA-Directed DNA Polymerase