Viral Load Analysis of Hepatitis C Virus in Huh7.5 Cell Culture System

AuthorRoghayeh Teimourpouren
AuthorZahra Meshkaten
AuthorAida Gholoubien
AuthorHosein Nomanien
AuthorSina Rostamien
OrcidZahra Meshkat [0000-0002-4032-7599]en
Issued Date2015-05-01en
AbstractBackground: Previous studies using cell culture systems for the replication of hepatitis C virus have opened new research dimensions, and paved the ways for further and detailed studies of the virus in vitro. Objectives: The purpose of the present study was to cultivate hepatitis C virus in a cell culture system and evaluate viral amplification. Materials and Methods: In order to propagate hepatitis C virus, cloned whole genome of virus, JFH-1, was used. JFH-1 cDNA was introduced into strain JM109 of Escherichia coli and plasmid, containing the viral genome was purified from transformed bacteria. After XbaI digestion, RNA synthesis was induced using T7 RNA polymerase enzyme. Next, eukaryotic cell line Huh 7.5 was transfected by the purified RNA. Finally, Huh-7.5 cell line was infected with replicated virus and viral load was determined using real-time PCR (Polymerase Chain Reaction). Results: The amount of viral load, which was measured using real-time PCR was 17592 IU/mL. Conclusions: In the present study, using cell culture, a high titer (in acceptable range) of infectious hepatitis C virus was produced. This method could be used in future studies.en
DOIhttps://doi.org/10.5812/jjm.8(5)2015.19279en
KeywordHepatitis C Virusen
KeywordDNAen
KeywordComplementaryen
KeywordJFH-1en
KeywordHuh 7.5en
KeywordReal-Time Polymerase Chain Reactionen
PublisherBrieflandsen
TitleViral Load Analysis of Hepatitis C Virus in Huh7.5 Cell Culture Systemen
TypeResearch Articleen

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