Martín J, Navas S, Quiroga J A, Colucci G, Pardo M, Carreño V
Department of Hepatology, Fundación Jiménez Díaz, Madrid, Spain.
J Med Virol. 1998 Apr;54(4):265-70. doi: 10.1002/(sici)1096-9071(199804)54:4<265::aid-jmv5>3.0.co;2-2.
Since the natural history of hepatitis C virus-associated liver disease and the therapeutic responsiveness might vary according to liver and blood mononuclear cells viral levels, it may be important to quantitate viral RNA in liver, blood mononuclear cells and serum, and to compare these data with genotype, biochemical and histologic data. A polymerase chain reaction-based assay available for serum hepatitis C virus RNA quantitation has been optimized to quantitate viral genomes in liver and peripheral blood mononuclear cells from 47 chronic hepatitis C patients. The procedure permitted hepatitis C virus RNA quantitation in freshly isolated mononuclear cells and in total RNA extracted from frozen mononuclear cells and liver tissue. The intrahepatic viral amount (median: 2.6 x 10(3) copies/microgram RNA; range: 0 to 3.6 x 10(4) copies/microgram RNA) correlated significantly with the hepatitis C virus RNA concentration in serum (r = 0.76, P < .001), but not in mononuclear cells. Viral RNA concentrations in liver (P < .001), serum (P < 0.01) and PBMC (P < 0.05) were significantly higher in hepatitis C virus genotype 1 patients (essentially type 1b) than in non-1 type cases, but were unrelated to biochemical or histologic indexes of disease activity. In conclusion, the optimized assay permit HCV RNA quantitation in liver and peripheral blood mononuclear cells, suggesting that serum viral level is an accurate measurement of intrahepatic viral burden.
由于丙型肝炎病毒相关肝病的自然病程以及治疗反应性可能因肝脏和血液单核细胞中的病毒水平而异,因此对肝脏、血液单核细胞和血清中的病毒RNA进行定量,并将这些数据与基因型、生化和组织学数据进行比较可能很重要。一种基于聚合酶链反应的血清丙型肝炎病毒RNA定量检测方法已得到优化,用于对47例慢性丙型肝炎患者肝脏和外周血单核细胞中的病毒基因组进行定量。该方法可对新鲜分离的单核细胞以及从冷冻单核细胞和肝组织中提取的总RNA中的丙型肝炎病毒RNA进行定量。肝内病毒量(中位数:2.6×10³拷贝/微克RNA;范围:0至3.6×10⁴拷贝/微克RNA)与血清中的丙型肝炎病毒RNA浓度显著相关(r = 0.76,P <.001),但与单核细胞中的病毒RNA浓度无关。丙型肝炎病毒1型患者(主要为1b型)肝脏(P <.001)、血清(P < 0.01)和外周血单个核细胞(PBMC)(P < 0.05)中的病毒RNA浓度显著高于非1型病例,但与疾病活动的生化或组织学指标无关。总之,优化后的检测方法可对肝脏和外周血单核细胞中的HCV RNA进行定量,表明血清病毒水平是肝内病毒载量的准确测量指标。