Further study of hepatitis C virus RNA detection in formalin-fixed, paraffin-embedded liver tissues by ligation-dependent polymerase chain reaction.
Ligation-dependent polymerase chain reaction (LD-PCR) is a novel, simple and sensitive PCR assay that could be useful for the molecular diagnosis of various viral infections. This assay uses two capture probes for RNA isolation and two hemiprobes for the subsequent PCR. To investigate its usefulness in the field of pathological diagnosis, further study of this new PCR technology to detect hepatitis C virus (HCV) RNA was carried out in formalin-fixed, paraffin-embedded (FFPE) liver tissues and the results compared with those obtained by the reverse transcription (RT)-PCR method. We analyzed routinely processed FFPE liver specimens of surgical origin from 13 anti-HCV-seropositive Japanese patients with hepatocellular carcinoma (HCC). As a result, HCV-RNA was detected in FFPE liver specimens in 12 of the tested patients (92.3%) by the LD-PCR method without the use of reverse transcription. In contrast, HCV-RNA was detectable in eight out of 13 cases (61.5%) by the RT-PCR method. These results indicated that LD-PCR was useful for HCV-RNA detection in routinely processed FFPE liver specimens.
Department of Pathology, National Institute of Infectious Diseases, Tokyo, Japan.
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Miyauchi
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Moriyama
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DY
Zhang
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K
Abe
D000328:Adult; D000368:Aged; D001483:Base Sequence; D006528:Carcinoma, Hepatocellular / Q000821:virology; D017931:DNA Primers / Q000737:chemistry; D015342:DNA Probes / Q000737:chemistry; D005557:Formaldehyde; D016174:Hepacivirus / Q000235:genetics* / Q000302:isolation & purification; D006526:Hepatitis C / Q000175:diagnosis / Q000821:virology*; D006801:Humans; D008099:Liver / Q000473:pathology / Q000821:virology*; D008113:Liver Neoplasms / Q000821:virology; D008875:Middle Aged; D008969:Molecular Sequence Data; D016612:Paraffin Embedding; D016133:Polymerase Chain Reaction / Q000379:methods*; D012367:RNA, Viral / Q000032:analysis* / Q000235:genetics; D016707:Tissue Fixation
D017931:DNA Primers; D015342:DNA Probes; D012367:RNA, Viral; D005557:Formaldehyde