Development of a reporter bovine viral diarrhea virus and initial evaluation of its application for high throughput antiviral drug screening

2012 ◽  
Vol 180 (1-2) ◽  
pp. 54-61 ◽  
Author(s):  
Zhen-Chuan Fan ◽  
R. Curtis Bird
Keyword(s):  
High Throughput ◽  
Drug Screening ◽  
Bovine Viral Diarrhea Virus ◽  
Antiviral Drug ◽  
Initial Evaluation ◽  
Bovine Viral Diarrhea ◽  
Diarrhea Virus ◽  
Viral Diarrhea ◽  
Viral Diarrhea Virus

scholarly journals Developing an Antiviral Drug Screening System for Anti-Bovine Viral Diarrhea Virus (BVDV) Therapies

2019 ◽  
Vol 69 (4) ◽  
pp. 1255
Author(s):  
A. Mokhtari ◽  
M. Mahzounieh ◽  
O. Madadgar ◽  
A. Ghalyanchi Langeroudi2
Keyword(s):  
Cell Line ◽  
Drug Screening ◽  
Bovine Viral Diarrhea Virus ◽  
Antiviral Drug ◽  
Bovine Viral Diarrhea ◽  
Screening System ◽  
Egfp Gene ◽  
Diarrhea Virus ◽  
Viral Diarrhea ◽  
Viral Diarrhea Virus

Bovine viral diarrhea virus (BVDV) is an economically important animal pathogen affecting cattle. Despite the use of vaccination, test and slaughter practices, BVD remains a serious problem of cattle breeding. This study was conducted in order to develop a cell line that expresses some of BVDV sub-replicons. BVDV-NADL NS3 and 5’UTR were cloned in pWPI-linker B lentiviral plasmid at the upstream of EGFP gene. Consequently, lentiviral vectors containing BVDV-NS3 and BVDV-5’UTR were produced by using the second-generation lentiviral packaging system. By these lentivectors, MDBK cells expressing BVDV-5’UTR and BVDV-NS3 partial fragments were prepared. The efficiency of the infection was evaluated by fluorescence microscopy, western blotting, and RT-PCR. The results indicated that the development of MDBK cell line expressing these transgenes provides a very sensitive antiviral drug screening system for anti-bovine viral diarrhea virus (BVDV) therapies.

scholarly journals Development of a Cell-Based High-Throughput Specificity Screen Using a Hepatitis C Virus-Bovine Viral Diarrhea Virus Dual Replicon Assay

2005 ◽  
Vol 49 (4) ◽  
pp. 1346-1353 ◽  
Author(s):  
Donald R. O'Boyle ◽  
Peter T. Nower ◽  
Julie A. Lemm ◽  
Lourdes Valera ◽  
Jin-Hua Sun ◽  
...  
Keyword(s):  
Hepatitis C Virus ◽  
Hepatitis C ◽  
High Throughput ◽  
Bovine Viral Diarrhea Virus ◽  
Bovine Viral Diarrhea ◽  
Diarrhea Virus ◽  
Viral Diarrhea ◽  
Reporter Enzyme ◽  
Ns3 Protease ◽  
Viral Diarrhea Virus

ABSTRACT The hepatitis C virus (HCV) replicon is a unique system for the development of a high-throughput screen (HTS), since the analysis of inhibitors requires the quantification of a decrease in a steady-state level of HCV RNA. HCV replicon replication is dependent on host cell factors, and any toxic effects may have a significant impact on HCV replicon replication. Therefore, determining the antiviral specificity of compounds presents a challenge for the identification of specific HCV inhibitors. Here we report the development of an HCV/bovine viral diarrhea virus (BVDV) dual replicon assay suitable for HTS to address these issues. The HCV reporter enzyme is the endogenous NS3 protease contained within the HCV genome, while the BVDV reporter enzyme is a luciferase enzyme engineered into the BVDV genome. The HTS uses a mixture of HCV and BVDV replicon cell lines placed in the same well of a 96-well plate and isolated in the same cell backgrounds (Huh-7). The format consists of three separate but compatible assays: the first quantitates the amount of cytotoxicity based upon the conversion of Alamar blue dye via cellular enzymes, while the second indirectly quantitates HCV replicon replication through measurement of the amount of NS3 protease activity present. The final assay measures the amount of luciferase activity present from the BVDV replicon cells, as an indicator of the specificity of the test compounds. This HCV/BVDV dual replicon assay provides a reliable format to determine the potency and specificity of HCV replicon inhibitors.

Prevention and elimination of bovine viral diarrhea virus infections in fetal fibroblast cells

2004 ◽  
Vol 64 (2) ◽  
pp. 113-118 ◽  
Author(s):  
M GIVENS ◽  
D STRINGFELLOW ◽  
C DYKSTRA ◽  
K RIDDELL ◽  
P GALIK ◽  
...  
Keyword(s):  
Bovine Viral Diarrhea Virus ◽  
Bovine Viral Diarrhea ◽  
Fibroblast Cells ◽  
Virus Infections ◽  
Fetal Fibroblast ◽  
Diarrhea Virus ◽  
Viral Diarrhea ◽  
Viral Diarrhea Virus

scholarly journals Identification and genotyping of a new subtype of bovine viral diarrhea virus 1 isolated from cattle with diarrhea

2021 ◽  
Vol 166 (4) ◽  
pp. 1259-1262
Author(s):  
Bin Tian ◽  
Dongjie Cai ◽  
Weiqiang Li ◽  
Qinglong Bu ◽  
Mingshu Wang ◽  
...  
Keyword(s):  
Bovine Viral Diarrhea Virus ◽  
Nasal Swab ◽  
Bovine Viral Diarrhea ◽  
Diarrhea Virus ◽  
Viral Diarrhea ◽  
Separate Branch ◽  
Genetic Subtype ◽  
Viral Diarrhea Virus ◽  
Cattle Farms ◽  
Subtype 1B

AbstractIn 2019, diarrhea cases occurred on cattle farms in Qionglai and Guang'an, Sichuan Province. Two out of 20 (10%) serum and nasal swab samples were positive when tested using a bovine viral diarrhea virus (BVDV) antigen-capture ELISA kit. Two non-cytopathic strains of BVDV were isolated and named QL1903 and GA190608, respectively. The nucleotide sequences of the genomes of the two isolates were 89.52% identical. Phylogenetic analysis based on the 5'-UTR sequence revealed that the BVDV isolate QL1903 belonged to BVDV subtype 1b, whereas isolate GA190608 clustered with strains HN1814, EN-19, and BJ09_26 in a separate branch, which has tentatively been classified as a new genetic subtype, "1v".

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