Viral load analysis of hepatitis C virus in Huh7.5 cell culture system

Teimourpour, R. and Meshkat, Z. and Gholoubi, A. and Nomani, H. and Rostami, S. (2015) Viral load analysis of hepatitis C virus in Huh7.5 cell culture system. Jundishapur Journal of Microbiology, 8 (5).

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Abstract

Background: 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. © 2015, Ahvaz Jundishapur University of Medical Sciences.

Item Type: Article
Additional Information: Cited By :5 Export Date: 16 February 2020 Correspondence Address: Meshkat, Z.; Antimicrobial Resistance Research Center, Mashhad University of Medical Sciences, P. O. Box: 9196773117, Iran
Uncontrolled Keywords: Complementary DNA Hepatitis C Virus Huh 7.5 JFH-1 Real-time polymerase chain reaction Article cell culture cell line Huh7 5 cell line nonhuman real time polymerase chain reaction virus genome virus infection virus load virus replication Escherichia coli Eukaryota
Subjects: WC Communicable Diseases
Divisions: Mashhad University of Medical Sciences
Depositing User: mr lib5 lib5
Date Deposited: 14 Mar 2020 07:28
Last Modified: 14 Mar 2020 07:28
URI: http://eprints.mums.ac.ir/id/eprint/16645

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