Cho S W, Hwang S G, Han D C, Jin S Y, Lee M S, Shim C S, Lee D W, Lee H B
Department of Internal Medicine, Soon Chun Hyang University Hospital, Seoul, Korea.
J Med Virol. 1996 Mar;48(3):227-33. doi: 10.1002/(SICI)1096-9071(199603)48:3<227::AID-JMV3>3.0.CO;2-A.
The cellular localization of hepatitis C virus (HCV) RNA in liver tissue was studied by nonisotopic in situ hybridization using a digoxigenin-labeled cDNA probe created during a polymerase chain reaction on samples from 16 patients with chronic HCV infection. Hybridization signals were recognized in the cytoplasm of the hepatocytes, and a few hepatocytes had hybridization signals in the nucleus as well. HCV RNA positive hepatocytes were found in 1 of 9 patients with chronic persistent hepatitis, 2 of 5 patients with chronic active hepatitis, and in each of 2 patients with chronic active hepatitis and cirrhosis. Positive signals were found in many hepatocytes within the lobule in liver sections of patients with advanced chronic active hepatitis. A number of HCV RNA positive hepatocytes were found in nodules, but not in the area of fibrosis. On the other hand, positive signals were found in a few hepatocytes scattered in the lobule in a patient with chronic persistent hepatitis. The mean ALT levels in the patients with positive signal (175.6 +/- 44.2 U/L) were significantly higher than in those without a signal (70.27 +/- 16.1 U/L) (P < 0.05). The findings suggest that a larger amount of HCV may be present during the advanced than during the early stages of type C hepatitis and nonisotopic in situ hybridization using a digoxigenin-labeled HCV cDNA probe created during a polymerase chain reaction deserves wider application for the detection of HCV replication in specimens.
采用地高辛素标记的cDNA探针，通过非同位素原位杂交技术，对16例慢性丙型肝炎病毒（HCV）感染患者的肝脏组织样本进行聚合酶链反应，研究HCV RNA的细胞定位。杂交信号在肝细胞的细胞质中被识别，少数肝细胞的细胞核中也有杂交信号。在9例慢性持续性肝炎患者中有1例发现HCV RNA阳性肝细胞，5例慢性活动性肝炎患者中有2例发现，2例慢性活动性肝炎合并肝硬化患者中均发现。在晚期慢性活动性肝炎患者的肝切片小叶内的许多肝细胞中发现了阳性信号。在结节中有许多HCV RNA阳性肝细胞，但在纤维化区域未发现。另一方面，在1例慢性持续性肝炎患者的小叶内散在的少数肝细胞中发现了阳性信号。有阳性信号的患者平均谷丙转氨酶（ALT）水平（175.6±44.2 U/L）显著高于无信号的患者（70.27±16.1 U/L）（P<0.05）。这些发现表明，丙型肝炎晚期可能比早期存在更多的HCV，并且使用聚合酶链反应产生的地高辛素标记的HCV cDNA探针进行非同位素原位杂交在检测标本中HCV复制方面值得更广泛应用。
