Recovery of duck hepatitis A virus 3 from a stable full-length infectious cDNA clone

Pan, Meng; Yang, Xiaorong; Du, Jige; Zhou, Lei; Ge, Xinna; Guo, Xin; Liu, Jinhua; Zhang, Dabing; Yang, Hanchun

doi:10.1016/j.virusres.2011.07.008

Cited by 7 publications

(5 citation statements)

References 28 publications

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“…They include duck hepatitis A virus genotypes 1 (DHAV-1; formerly duck hepatitis virus type 1) [2], 2 (DHAV-2) [3], and 3 (DHAV-3) [4], members of the species Avihepatovirus A of the genus Avihepatovirus in the family Picornaviridae [5] (http://www.picornaviridae.com/avihepatovirus/avihepatovirus.htm), and duck hepatitis virus type 2 (DHV-2) [6–8] and duck hepatitis virus type 3 (DHV-3) [9], which are currently classified within the genus Astrovirus in the family Astroviridae [10, 11]. Compared with DHV-2 and DHV-3, the three DHAV genotypes can cause more severe diseases [1, 3, 12, 13]. Among the DHAV genotypes, DHAV-1 is known to be worldwide in distribution [1]; DHAV-2 has only been reported in Taiwan, China [3]; and DHAV-3 has been found in South Korea [4], mainland China [14], and Vietnam [15].…”

Section: Introductionmentioning

confidence: 99%

“…The DHAV polyprotein appears to be cleaved into three structural (VP0, VP3 and VP1) and 8–9 nonstructural (2A1, 2A2 [or 2A2, 2A3], 2B, 2C, 3A, 3B, 3C and 3D) proteins [3, 4, 19–21]. Comparative sequence analysis demonstrates that DHAV-3 strains share low identity at the nucleotide (genome: 70–73%) and amino acid (polyprotein: 82–83%) level with DHAV-1 strains [4, 12, 22]. Moreover, DHAV-3 and DHAV-1 differ greatly in their genome lengths [4, 12, 19–21].…”

Section: Introductionmentioning

confidence: 99%

“…Comparative sequence analysis demonstrates that DHAV-3 strains share low identity at the nucleotide (genome: 70–73%) and amino acid (polyprotein: 82–83%) level with DHAV-1 strains [4, 12, 22]. Moreover, DHAV-3 and DHAV-1 differ greatly in their genome lengths [4, 12, 19–21]. Previous works have shown that DHAV-3 has no common antigens with DHAV-1 in virus neutralization tests [4, 14].…”

Section: Introductionmentioning

confidence: 99%

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Effect of fetal calf serum on propagation of duck hepatitis A virus genotype 3 in duck embryo fibroblast cells

Wang

Liu

et al. 2019

BMC Vet Res

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Background Duck viral hepatitis (DVH) is a highly contagious viral disease affecting ducks. It can be caused by five agents, including duck hepatitis A virus genotypes 1 (DHAV-1), 2 (DHAV-2), and 3 (DHAV-3), as well as duck hepatitis virus 2 and duck hepatitis virus 3. Since 2007, DHAV-3 has been known to be the most prevalent in East and South Asia. So far, the information regarding the propagation of DHAV-3 in cultured cells is limited. In this study, we describe the comparative studies on the growth properties of DHAV-3 in primary duck embryo fibroblast (DEF) cells using two different strains: a virulent strain C-GY and an attenuated strain YDF120. The effect of fetal calf serum (FCS) and chick serum (CS) on DHAV-3 replication and the mechanism of the inhibitory effect conferred by FCS were also investigated. Results Following serial passages, both C-GY and YDF120 failed to produce cytopathic effect and plaques. The combined quantitative real-time PCR and indirect immunofluorescence staining methods showed that the two viruses could be propagated productively in DEF cells. Investigation of the viral growth kinetics revealed that the two viruses replicated in DEF cells with similar efficiencies, while the viral load of the virulent C-GY strain peaked more rapidly when compared with the attenuated YDF120 strain. Neutralization assay and time-of-drug-addition study indicated that FCS displayed inhibitory effect on DHAV-3 replication. Analysis on the mechanism of action of FCS against DHAV-3 demonstrated that the inhibitory effect was reflected at three steps of the DHAV-3 life cycle including adsorption, replication, and release. Conclusions Both virulent and attenuated DAHV-3 strains can establish noncytocidal, productive infections in DEF cells. The virulent strain replicates more rapidly than the attenuated strain in early infection period. FCS has an inhibitory effect on DHAV-3 replication, which may be attributed to action of a non-specific inhibitory factor present in FCS directly on the virus. These findings may provide new insights into the development of potential antiviral agents.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Effect of fetal calf serum on propagation of duck hepatitis A virus genotype 3 in duck embryo fibroblast cells

Wang

Liu

et al. 2019

BMC Vet Res

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“…Reverse genetics system is particularly useful for RNA viruses since RNA genomes are difficult to manipulate directly. Although the previous research has reported about the establishment of the RNA-launched infectious clone of DHAV-1 [ 23 ] and DHAV-3 [ 24 ], the in vitro transcription will raise the operation difficulty and experiment cost. The DNA-launched infectious system possessed self-cleaving ribozyme elements at both termini of the viral genomic cDNA, which was expected to retain the authentic terminal nucleotide sequences of the viral genome [ 28 ].…”

Section: Discussionmentioning

confidence: 99%

“…Up to date, there are at least four types of viral RNA rescued system, including RNA-launched system [ 18 ], helper virus-launched system [ 19 ] and two DNA-launched systems based on cellular RNA polymerases I and II [ 20 – 22 ]. Although the infectious DHAV genomic RNA could be harvested in vitro from a full-length DHAV cDNA clone by the RNA-launched reverse genetics system [ 23 , 24 ], the operative strategy still depends on in vitro transcription. Compared to the RNA-launched infectious clone, the RNA polymerases II-based DNA-launched infectious system is able to generate homogenous RNA transcripts from transfected cDNA clone in vivo , allowing for higher rescue efficiency with less cost and labor by skipping in vitro RNA transcription [ 25 – 27 ].…”

Section: Introductionmentioning

confidence: 99%

Construction and characterization of an improved DNA-launched infectious clone of duck hepatitis a virus type 1

Chen¹,

Zhang²,

Lin³

et al. 2017

Virol J

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BackgroundDNA-launched infectious system is a useful tool with high rescue efficiency that allows the introduction of mutations in specific positions to investigate the function of an individual viral element. Rescued virus particles could be harvested by directly transfecting the DNA-launched recombinant plasmid to the host cells, which will reduce labor and experimental cost by skipping the in vitro transcription assay.MethodsA total of four overlapping fragments covering the entire viral genome were amplified and then were assembled into a transformation vector based on pIRES2-EGFP to establish the DNA-launched infectious system of duck hepatitis A virus type 1 (DHAV-1), named pIR-DHAV-1. Reverse transcription polymerase chain reaction (RT-PCR) detection, quantitative real-time polymerase chain reaction (qRT-PCR), western blotting assay and indirect immunofluorescence (IFA) were conducted for rescued virus identification. A total of 4.0 μg of recombinant plasmid of pIR-DHAV-1 and in vitro transcribed product of 4.0 μg of RNA-launched infectious clone named pR-DHAV-1 were transfected into BHK-21 cells to analyze the rescue efficiency. Following that, tissue tropism of rescued virus (rDHAV-1) and parental virus (pDHAV-1) were assayed for virulence testing in 1-day-old ducklings.ResultsRescued virus particles carry the designed genetic marker which could be harvested by directly transfecting pIR-DHAV-1 to BHK-21 cells. The qRT-PCR and western blotting results indicated that rDHAV-1 shared similar growth characteristics with pDHAV-1. Furthermore, DNA-launched infectious system possessed much higher rescue efficiency assay compared to RNA-launched infectious system. The mutation at position 3042 from T to C has no impact on viral replication and tissue tropism. From 1 h post infection (hpi) to 48 hpi, the viral RNA copies of rDHAV-1 in liver were the highest among the six tested tissues (with an exception of thymus at 6 hpi), while the viral RNA copy numbers in heart and kidney were alternately the lowest.ConclusionWe have constructed a genetically stable and highly pathogenic DNA-launched infectious clone, from which the rescued virus could be harvested by direct transfection with recombinant plasmids. rDHAV-1 shared similar growth characteristics and tissue tropism with pDHAV-1. The DNA-launched infectious system of DHAV-1 possessed higher rescue efficiency compared to the traditional RNA-launched infectious system.

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Advances in the Duck Hepatitis A virus and lessons learned from those in recent years

Zhao,

Wu,

Wang

et al. 2024

Microbial Pathogenesis

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Recovery of duck hepatitis A virus 3 from a stable full-length infectious cDNA clone

Cited by 7 publications

References 28 publications

Effect of fetal calf serum on propagation of duck hepatitis A virus genotype 3 in duck embryo fibroblast cells

Effect of fetal calf serum on propagation of duck hepatitis A virus genotype 3 in duck embryo fibroblast cells

Construction and characterization of an improved DNA-launched infectious clone of duck hepatitis a virus type 1

Advances in the Duck Hepatitis A virus and lessons learned from those in recent years

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