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

采用高灵敏度原位杂交技术（ISH），结合na修饰的寡核苷酸探针和无生物素的酪胺扩增系统（CSAII），成功地检测了福尔马林固定石蜡包埋的肝脏切片中HCV-RNA的含量。在嵌合小鼠和11例HCV感染患者中9例（82%）的HCV感染肝组织中均证实了HCV- rna。HCV-RNA未被鉴定为慢性乙型肝炎、脂肪肝、自身免疫性肝炎和肝细胞癌。巢式RT-PCR证实了11例肝脏中8例（73%）的基因型（1b型6个病变，2a型2个病变）。巢式RT-PCR可用于HCV基因分型检测，而高灵敏度的ISH技术在HCV基因型之间发生交叉反应。

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.