Detection of HPV‐16 in cell lines and cervical lavage specimens by a polymerase chain reaction‐enzyme immunoassay assay

François Coutlée, Linda Bobo, Hawwari Abbass, Gina Dalabetta, Ned E. Hook, Keerti Shah, Raphael P. Viscidi

Research output: Contribution to journalArticlepeer-review

Abstract

A gene amplification method that combines the polymerase chain reaction with detection of amplified DNA in a solution hybridization/enzyme immunoassay (PCR‐EIA) was developed for HPV‐16 DNA. Samples were amplified with primers for the E7‐E1 region of HPV‐16. Amplified DNA products were identified and quantitated by hybridization in solution with a biotinylated RNA probe. Labeled DNA/RNA hybrids were measured semiquantitatively in an enzyme immunoassay using solid phase anti‐biotin antibody and liquid phase B‐d‐galactosidase labeled monoclonal antibody against DNA‐RNA hybrids. Enzyme bound to the solid phase was quantitated with a fluorogenic substrate. The assay was linear over 2 log10 dilutions of SiHa cells and the detection limit was three copies of HPV‐16 genome. The sensitivity of PCR‐EIA for detection of PCR amplified products compared favorably with slot and Southern blots using a 32P‐labeled RNA probe. The assay was used to assess HPV‐16 infection of uterine cervix in women attending a clinic for sexually transmitted diseases. Twenty‐one of the 81 specimens (25.9%), obtained by cervicovaginal lavage, were positive for HPV‐16 by PCR‐EIA. The assay provides a convenient means to objectively measure HPV DNA amplified with PCR. © 1992 Wiley‐Liss, Inc.

Original languageEnglish (US)
Pages (from-to)22-29
Number of pages8
JournalJournal of Medical Virology
Volume37
Issue number1
DOIs
StatePublished - May 1992

Keywords

  • HPV
  • PCR
  • biotin
  • genital cancer
  • nonisotopic

ASJC Scopus subject areas

  • Virology
  • Infectious Diseases

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