Castillo I, Bartolomé J, Quiroga J A, Carreño V
Department of Gastroenterology, Fundación Jiménez Díaz, Madrid, Spain.
J Virol Methods. 1992 Jul;38(1):71-9. doi: 10.1016/0166-0934(92)90170-i.
Different methods for the isolation of hepatitis C virus RNA and several sets of primers from the 5' untranslated, core, NS3, NS3/NS4 and NS5 regions of the hepatitis C virus genome were used for the detection of the viral genome by the polymerase chain reaction. Serum samples from 10 patients with chronic hepatitis C and 10 healthy controls were studied. The best method for the RNA extraction was with cold guanidinium isothiocyanate followed by a denaturation step prior to the polymerase chain reaction. Using this method, the percentage of positivity to hepatitis C virus sequences in serum depending on the region amplified were: 5' untranslated, 90%; Core, 20%; NS3, 80%; NS3/NS4, 60% and NS5, 60%.
采用不同方法从丙型肝炎病毒基因组的5'非编码区、核心区、NS3区、NS3/NS4区和NS5区分离丙型肝炎病毒RNA及几组引物，通过聚合酶链反应检测病毒基因组。研究了10例慢性丙型肝炎患者和10例健康对照者的血清样本。RNA提取的最佳方法是使用冷异硫氰酸胍，然后在聚合酶链反应之前进行变性步骤。使用该方法，血清中丙型肝炎病毒序列的阳性率根据扩增区域不同分别为：5'非编码区90%；核心区20%；NS3区80%；NS3/NS4区60%；NS5区60%。
