Detection of hepatitis C virus RNA sequences by polymerase chain reaction in fixed liver tissue.
Currently, the most reliable method for the diagnosis of hepatitis C virus (HCV) infection is the detection of viral sequences by the reverse transcription double polymerase chain reaction (RT/PCR) in serum or liver samples. We demonstrate here that noncoding region sequences (NT) of HCV were amplifiable by RT/PCR in guanidinium extracts of formalin-fixed (for 6 to 48 h), paraffin-embedded liver sections of patients with chronic hepatitis C. In contrast, core and nonstructural region sequences of HCV were not detectable in fixed tissues by PCR amplification. Boiling of routinely processed tissue sections in water containing Chelex-100, a method for extraction of amplifiable hepatitis B virus DNA, was not successful. The amount of nucleic acid extracts from fixed liver sections needed for amplification of NT region sequences was over 1000 times larger than that of extracts from frozen tissue. This method will be useful for diagnostic and investigative studies of HCV infection.
Department of Pathology and Laboratory Medicine, Tulane University School of Medicine, New Orleans, Louisiana.
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G
Akyol
S
S
Dash
Y S
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Shieh
J S
JS
Malter
M A
MA
Gerber
D000368:Aged; D001483:Base Sequence; D017931:DNA Primers / Q000737:chemistry; D005260:Female; D016174:Hepacivirus / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D008099:Liver / Q000382:microbiology*; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes / Q000737:chemistry; D016612:Paraffin Embedding; D016133:Polymerase Chain Reaction; D012367:RNA, Viral / Q000302:isolation & purification*; D016707:Tissue Fixation
D017931:DNA Primers; D015345:Oligonucleotide Probes; D012367:RNA, Viral