Improved detection of viral RNA isolated from liquid-based cytology samples

Mol Diagn. 2001 Jun;6(2):125-30. doi: 10.1054/modi.2001.25320.


Background: Molecular diagnosis requires the ability to obtain high-quality nucleic acids that are representative of the disease state. We evaluated the recovery and detection of limiting amounts of viral oncogenic RNA from cells fixed in liquid-based cytology media.

Methods and results: Serial dilutions of a human papillomavirus (HPV)-positive cell line fixed in a liquid media was used as a model system. Total nucleic acid (TNA) extraction produced RNA with clearly visible ribosomal bands even after one year of storage. These TNA extracts, treated with DNase-I, were used in an RT-PCR assay for HPV-16 E6-E7 oncogenic transcripts. With chemiluminscent Southern blot detection, samples with one HPV-positive cell in 30,000 were consistently detected.

Conclusion: PreservCyt-fixed cells can yield RNA suitable for molecular assays even after one year of storage.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Blotting, Southern
  • Chemistry, Clinical / methods*
  • DNA / analysis*
  • Electrophoresis, Agar Gel
  • Female
  • Humans
  • Luminescent Measurements
  • Molecular Epidemiology
  • Oncogene Proteins, Viral / biosynthesis
  • Papillomaviridae / genetics
  • Papillomavirus E7 Proteins
  • RNA, Viral / analysis*
  • Repressor Proteins*
  • Reverse Transcriptase Polymerase Chain Reaction
  • Sensitivity and Specificity
  • Tumor Cells, Cultured
  • Uterine Cervical Neoplasms / virology


  • E6 protein, Human papillomavirus type 16
  • Oncogene Proteins, Viral
  • Papillomavirus E7 Proteins
  • RNA, Viral
  • Repressor Proteins
  • oncogene protein E7, Human papillomavirus type 16
  • DNA