Detection of human papillomavirus type-16 DNA utilising microtitre-plate based amplification reactions and a solid-phase enzyme-immunoassay detection system

S Cavuslu, W G Starkey, J N Kaye, C Biswas, C Mant, B Kell, P Rice, J M Best, J Cason

Research output: Contribution to journalArticlepeer-review

16 Citations (Scopus)

Abstract

The development of a nested polymerase chain reaction (PCR) assay to detect low concentrations of human papillomavirus type-16 (HPV-16) DNA for epidemiological studies is described. The PCR utilises primers located in the E5 open reading frame, has an analytical sensitivity of 4 HPV-16 genomes and does not produce amplicons from other common genital HPVs (types-6, -11, -18, -31 and 33). This assay was carried out in 96-well plates utilising internal primers labelled with dinitrophenol (DNP) and biotin so that amplicons can be captured onto streptavidincoated plates and detected using an alkaline phosphatase-labelled monoclonal antibody to DNP. The assay was effective for detecting HPV-16 DNA in plasmids, cell-lines and, both freshly collected or archival (formalin-fixed/paraffin embedded) clinical specimens. This system is therefore suitable for epidemiological studies to identify individuals infected with HPV-16 DNA in episomal form who may be at increased risk of developing anogenital carcinomas.

Original languageEnglish
Pages (from-to)59-69
Number of pages11
JournalJournal of Virological Methods
Volume58
Issue number1-2
DOIs
Publication statusPublished - 26 Apr 1996

Keywords

  • Cervical Intraepithelial Neoplasia/pathology
  • DNA Primers
  • DNA, Viral/analysis
  • Evaluation Studies as Topic
  • Female
  • HeLa Cells
  • Humans
  • Immunoenzyme Techniques
  • Oncogene Proteins, Viral/genetics
  • Papillomaviridae/genetics
  • Papillomavirus E7 Proteins
  • Polymerase Chain Reaction/methods
  • Repressor Proteins
  • Sensitivity and Specificity
  • Tumor Cells, Cultured
  • Uterine Cervical Neoplasms/pathology

Fingerprint

Dive into the research topics of 'Detection of human papillomavirus type-16 DNA utilising microtitre-plate based amplification reactions and a solid-phase enzyme-immunoassay detection system'. Together they form a unique fingerprint.

Cite this