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Polymerase-chain-reaction-based semi-quantification of hepatitis D viraemia in patients treated with high doses of alpha 2b interferon.
- Source :
-
Research in virology [Res Virol] 1994 Sep-Oct; Vol. 145 (5), pp. 287-95. - Publication Year :
- 1994
-
Abstract
- To study the antiviral efficacy of high doses of alpha 2b interferon (alpha 2b-IFN) for chronic hepatitis D treatment, we used polymerase-chain-reaction(PCR)-based semi-quantitative detection of HDV RNA. The semi-quantification method used was based on the appearance of a positive amplification signal as a function of the number of PCR cycles. By amplifying dilutions (10(-1)-10(-8)) of an HDV-positive woodchuck liver RNA, we confirmed that exponential amplification efficacy occurred at between 20 and 30 cycles. Positive signals were observed from dilution 10(-2) (gel electrophoresis after 20 cycles of PCR) to dilution 10(-7) (hybridization after 30 cycles of PCR). To characterize the HDV RNA level in sera of 8 patients treated with alpha 2b-IFN (10 MU/3 times a week) for 1 year, we extracted RNA from serum samples taken every 6 months. All samples were amplified in parallel for 20 and 30 PCR cycles. Analysis of HDV cDNA after ethidium bromide/agarose gel electrophoresis and after molecular hybridization (100 times more sensitive than gel analysis), enabled us to grade the signals observed from negative to positive as 1+, 2+, 3+ and 4+, with all results being positive. Three types of evolution of HDV viraemia were evidenced among the 8 treated patients. HDV replication continued to occur at a high level at the 6th and 12th month in 2 patient sera. For 2 other patients, an HDV RNA decrease or disappearance was evidenced in the serum at the 6th month; however, viral replication recurred at a higher level at the 12th month.(ABSTRACT TRUNCATED AT 250 WORDS)
- Subjects :
- Adult
Base Sequence
Female
Hepatitis D complications
Hepatitis Delta Virus drug effects
Hepatitis, Chronic drug therapy
Humans
Interferon alpha-2
Liver Cirrhosis complications
Male
Molecular Sequence Data
Recombinant Proteins
Virus Replication drug effects
Hepatitis D drug therapy
Interferon-alpha therapeutic use
Polymerase Chain Reaction methods
RNA, Viral blood
Viremia drug therapy
Subjects
Details
- Language :
- English
- ISSN :
- 0923-2516
- Volume :
- 145
- Issue :
- 5
- Database :
- MEDLINE
- Journal :
- Research in virology
- Publication Type :
- Academic Journal
- Accession number :
- 7839006
- Full Text :
- https://doi.org/10.1016/s0923-2516(07)80033-1