Detection and partial sequencing of hepatitis C virus RNA in the liver.
Detection and partial sequencing of hepatitis C virus RNA in the liver.
复制标题
肝脏中丙型肝炎病毒RNA的检测和部分测序。
DOI：
10.1016/0016-5085(91)90537-u
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发表时间：
1991
期刊：
影响因子：
29.4
通讯作者：
M. Ohto
中科院分区：
文献类型：
--
作者：
K. Hosoda;O. Yokosuka;M. Omata;N. Kato;M. Ohto
为检测丙型肝炎病毒RNA，从肝组织中提取总RNA，反转录为互补DNA，聚合酶链式反应扩增。反应产物用溴化乙锭在丙烯酰胺凝胶中染色，并与放射性标记探针杂交。因此，在27例来自非甲非乙型慢性肝病患者的肝组织标本中有17例(63%)检测到丙型肝炎病毒RNA。在这27例患者中，抗丙型肝炎病毒阳性患者的17例肝组织中有12例(71%)检出病毒RNA，而在10例抗丙型肝炎病毒阴性患者的肝组织中检出5例(50%)病毒RNA。对17个RNA阳性样本进行扩增的互补DNA(35个核苷酸)的直接测序显示，与报道的丙型肝炎病毒互补DNA序列只有66%至77%的同源性。这些结果表明，目前大多数抗丙型肝炎病毒阳性患者感染了丙型肝炎病毒，部分抗丙型肝炎病毒阴性非甲、非乙型肝炎患者肝脏中存在丙型肝炎病毒。丙型肝炎病毒RNA的检测似乎为丙型肝炎病毒感染的研究提供了一个有用的指标。
To detect hepatitis C virus RNA, total RNA was extracted from liver tissue, reverse transcribed to complementary DNA, and amplified by polymerase chain reaction. The reaction products were analyzed by ethidium bromide staining in acrylamide gel and hybridization with a radiolabeled probe. Hepatitis C virus RNA was thereby detected in 17 of 27 (63%) liver tissue specimens obtained from patients with non-A, non-B chronic liver diseases. Of these 27 patients, viral RNA was detected in 12 of 17 (71%) liver tissues from anti-hepatitis C virus-positive patients and in 5 of 10 (50%) liver tissues from anti-hepatitis C virus-negative patients. Direct sequencing of amplified complementary DNA (35 nucleotides) of the 17 RNA-positive samples showed only 66% to 77% homology to the reported hepatitis C virus complementary DNA sequence. These results indicate that the majority of anti-hepatitis C virus-positive patients are currently infected with hepatitis C virus, and some of the anti-hepatitis C virus-negative patients with non-A, non-B hepatitis are harboring hepatitis C virus in the liver. Detection of hepatitis C virus RNA appears to provide a useful indicator in the study of hepatitis C virus infection.