Proteomics reveals the effect of type I interferon on the pathogenicity of duck hepatitis A virus genotype 3 in Pekin ducks

The co-circulation of duck hepatitis A virus subtypes 1 (DHAV-1) and 3 (DHAV-3) in ducklings has resulted in significant economic losses. Because ducklings infected with DHAV-1 or DHAV-3 show similar clinical signs and gross lesions, it is important to discriminate these subtypes as early as possible for better clinical management. On the basis of multiple alignments of the 5′-noncoding region sequences of strains DHAV-1 and DHAV-3, universal and type-specific primers were designed and synthesized. Using the primers in a one-tube reverse transcription-PCR (RT-PCR) assay, reference strains of DHAV-1 and DHAV-3 (isolated over a span of 60 years and covering many different countries) were successfully amplified, indicating that the primer sequences were completely conserved. The amplicon sequences results and the sizes of amplicons from reference DHAV-1 and DHAV-3 isolates correlated completely with their genotypes. Moreover, with this one-tube RT-PCR system, the amplicon sizes of liver samples of reference DHAV-1- or DHAV-3-infected birds matched perfectly with their respective genotypes, as determined by virus isolation and neutralization tests. No other RNA viruses of duck origin were detected with the synthesized primers. The sensitivity of viral RNA detection was 10 pg. With this system, 20% genotype 1, 45% genotype 3, and 9% co-infection of the two genotypes were detected in 55 clinical samples. This novel approach could be used for the rapid genotyping DHAV-1 and/or DHAV-3 infection in routine clinical surveillance or epidemiologic screening.

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Comparative analysis of virus-host interactions caused by a virulent and an attenuated duck hepatitis A virus genotype 1

PLoS ONE ◽

10.1371/journal.pone.0178993 ◽

2017 ◽

Vol 12 (6) ◽

pp. e0178993 ◽

Cited By ~ 15

Author(s):

Xumin Ou ◽

Sai Mao ◽

Jingyu Cao ◽

Anchun Cheng ◽

Mingshu Wang ◽

...

Keyword(s):

Comparative Analysis ◽

Hepatitis A ◽

Hepatitis A Virus ◽

Genotype 1 ◽

Virus Genotype ◽

Host Interactions ◽

Duck Hepatitis

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Fetal Calf Serum Exerts an Inhibitory Effect on Replication of Duck Hepatitis A Virus Genotype 1 in Duck Embryo Fibroblast Cells

Viruses ◽

10.3390/v12010080 ◽

2020 ◽

Vol 12 (1) ◽

pp. 80

Author(s):

Minghang Wang ◽

Lili Chai ◽

Suyun Liang ◽

Junfeng Lv ◽

Lixin Yang ◽

...

Keyword(s):

Hepatitis A ◽

Hepatitis A Virus ◽

Calf Serum ◽

Plaque Formation ◽

Genotype 1 ◽

Duck Embryo Fibroblast ◽

Virus Genotype ◽

Extracellular Virus ◽

Duck Hepatitis ◽

Duck Embryo

Among the causative agents of duck viral hepatitis, duck hepatitis A virus genotype 1 (DHAV-1) is the most common virus reported in most outbreaks worldwide. How to propagate DHAV-1 in cell cultures efficiently remains a problem to be explored. Here, we aimed to test the effect of serum type on DHAV-1 replication in duck embryo fibroblast (DEF) cells. Comparative studies involved virus culture and passage, observation of cytopathic effect (CPE), virus quantification, and plaque formation assay. From the results of these investigations, we conclude that use of chicken serum (CS) in maintenance medium allows DHAV-1 to establish productive, cytocidal infection in DEF cells, whereas FCS exerts inhibitory effects on DHAV-1 replication, CPE development, and plaque formation. By using a neutralization test, we found that the direct action of FCS on virions is likely to play a key role in inhibiting DHAV-1 replication in DEF cells. Mechanism analyses revealed that FCS inhibits DHAV-1 replication at virus adsorption and reduces extracellular virus yields. The present work may shed light on a new perspective for antiviral agent development, and have provided a virus–host cell system for further studies on molecular mechanism involved DHAV-1 replication and pathogenesis.

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