Journal
AMERICAN JOURNAL OF CLINICAL PATHOLOGY
Volume 119, Issue 1, Pages 95-100Publisher
OXFORD UNIV PRESS INC
DOI: 10.1309/33TAJLB748KLMXVG
Keywords
hepatitis C virus; lymphocyte; complementary DNA; PCR; polymerase chain reaction; nested PCR; DNA sequencing
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We established a cell culture system for the replication of hepatitis C virus (HCV) by using human T and B leukemia cell lines. These 2 cell lines were infected in vitro by using HCV-positive pooled patient serum samples. HCV RNA was extracted from infected cell lines at different times after infection, and a sequence of the virus 5' untranslated region was analyzed. Hepatitis C minus-strand RNA was detected in the infected cell lines by highly strand-specific rTth (recombinant Thermus thermophilus DNA polymerase)based reverse transcription followed by a novel, highly sensitive, single-tube nested polymerase chain reaction (PCR) method. PCR products were analyzed by direct DNA sequencing. These results indicate that the HCV can replicate in T and B lymphocytes. This model should represent a valuable tool for the detailed study of the initial steps of the HCV replication cycle and for the evaluation of antiviral molecules.
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