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      An immunoassay for specific amplified HCV sequences.

      Journal of Virological Methods

      Hepatitis, Chronic, Adult, Animals, Antibodies, Monoclonal, Base Sequence, Blotting, Southern, DNA, Viral, analysis, Hepacivirus, isolation & purification, Hepatitis C, diagnosis, Humans, Immunoassay, methods, Liver, microbiology, Mice, Molecular Sequence Data, Polymerase Chain Reaction, RNA, Viral, blood

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          Abstract

          The direct detection of viraemia could improve greatly the efficacy of presently available assays. Due to its sensitivity, the polymerase chain reaction represents the method of choice for direct detection of viral nucleic acid. However, the clinical application of this method is hampered by the requirement of hybridization with radioactively labelled probes. In this study we demonstrate that HCV cDNA, amplified by the polymerase chain reaction from both liver tissues and sera, can be detected specifically by a new non-radioisotopic method, DNA enzyme immunoassay, that is based on an antibody that selectively recognizes double, but not single-stranded DNA. The assay reveals the hybridization events, independently from the DNA sequences, and therefore can be used with any combination of primers and probes. Most importantly, the method has a conventional ELISA format and is compatible with standard facilities of clinical laboratories. The availability of this new approach for revealing amplified sequences may facilitate greatly the use of PCR as the method of choice for early diagnosis of HCV infection.

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          1660488

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