Identification of genotype of hepatitis C virus using high-sensitivity in situ hybridization

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We were successful to detect HCV-RNA in formalin-fixed, paraffin-embedded sections of liver by using high-sensitivity in situ hybridization (ISH) combined with LNA-modified oligonucleotide probe and biotin-free tyramide amplification system (CSAII). HCV-RNA was demonstrated in HCV-infected liver tissues from both chimeric mice and 9 (82%) of 11 patients with HCV infection. HCV-RNA was not identified chronic hepatitis B, fatty liver, autoimmune hepatitis, and hepatocellular carcinoma. Nested RT-PCR confirmed the genotype in 8 (73%) of 11 liver (type 1b: 6 lesions and type 2a: 2 lesions). HCV genotyping could be detected by nested RT-PCR, while high-sensitivity ISH technique occurred cross reaction among the HCV genotypes.