HCV RNA levels in hepatocellular carcinomas and adjacent non-tumorous livers

Srikanta Dash, Romil Saxena, Jane Myung, Tanya Rege, Hideyuki Tsuji, Paul Gaglio, Robert F. Garry, Swan N. Thung, Michael A. Gerber

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

To determine the antiviral effects of drugs targeted to hepatitis C virus (HCV) in chronic hepatitis patients, an accurate quantitative method with high sensitivity is needed. Reverse transcription nested polymerase chain reaction (RT-PCR) is the most sensitive method for the detection of HCV sequences in clinical specimens. However, this method is not quantitative. For this purpose, a quantitative competitive assay was developed that combines RT and PCR followed by image analysis to quantify HCV RNA. This assay targets the highly conserved 5' non-coding region of HCV and is based on the co-amplification of wild type HCV RNA with known amounts of mutant synthetic RNA. The mutant internal control used in these experiments differs from the wild type RNA by two nucleotide substitutions, which introduces an internal restriction enzyme site. In this report, this method was used to determine the levels of positive strand RNA in 11 HCV positive hepatocellular carcinomas (HCC) and compared these with adjacent non-tumorous liver tissue. To confirm that the difference in viral titers is not related to variations in the amount of RNA used in the assay, glyceraldehyde 3-phosphate dehydrogenase (GAPDH) mRNA was also assessed by competitive RT-PCR in all tissue extracts. Using this competitive assay it was determined that HCV RNA levels in the liver and tumor samples ranged from 103 to 106 molecules per μg of total RNA which is similar to previous reports. Interestingly, the amount of HCV in all the non-tumorous liver specimens were found to be significantly higher (P

Original languageEnglish (US)
Pages (from-to)15-23
Number of pages9
JournalJournal of Virological Methods
Volume90
Issue number1
DOIs
StatePublished - Oct 2000
Externally publishedYes

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Hepacivirus
Hepatocellular Carcinoma
RNA
Liver
Polymerase Chain Reaction
Reverse Transcription
Glyceraldehyde-3-Phosphate Dehydrogenases
Tissue Extracts
Chronic Hepatitis
Antiviral Agents
Nucleotides
Messenger RNA
Enzymes
Neoplasms

Keywords

  • Hepatitis C virus
  • Non-tumorous liver
  • RNA

ASJC Scopus subject areas

  • Virology

Cite this

HCV RNA levels in hepatocellular carcinomas and adjacent non-tumorous livers. / Dash, Srikanta; Saxena, Romil; Myung, Jane; Rege, Tanya; Tsuji, Hideyuki; Gaglio, Paul; Garry, Robert F.; Thung, Swan N.; Gerber, Michael A.

In: Journal of Virological Methods, Vol. 90, No. 1, 10.2000, p. 15-23.

Research output: Contribution to journalArticle

Dash, S, Saxena, R, Myung, J, Rege, T, Tsuji, H, Gaglio, P, Garry, RF, Thung, SN & Gerber, MA 2000, 'HCV RNA levels in hepatocellular carcinomas and adjacent non-tumorous livers', Journal of Virological Methods, vol. 90, no. 1, pp. 15-23. https://doi.org/10.1016/S0166-0934(00)00199-3
Dash, Srikanta ; Saxena, Romil ; Myung, Jane ; Rege, Tanya ; Tsuji, Hideyuki ; Gaglio, Paul ; Garry, Robert F. ; Thung, Swan N. ; Gerber, Michael A. / HCV RNA levels in hepatocellular carcinomas and adjacent non-tumorous livers. In: Journal of Virological Methods. 2000 ; Vol. 90, No. 1. pp. 15-23.
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