Comparative proteomic analysis revealed complex responses to classical/novel duck reovirus infections in Cairna moschata

Yun, Tao; Hua, Jionggang; Ye, Weicheng; Yu, Bin; Chen, Liu; Zheng, Na; Zhang, Cun

doi:10.1038/s41598-018-28499-3

Cited by 13 publications

(35 citation statements)

References 37 publications

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“…In China, NDRV has emerged in recent years and is a current common genotype [13,16]. Recently, a group of newly emerging DRVs [24][25][26] was confirmed and characterized in Cherry Valley duck in China [18]. The NDRV from the mainly infected ducklings caused and other avian reovirus strains based on the σC gene in the phylogenetic tree.…”

Section: Discussionmentioning

confidence: 99%

A TaqMan-based real-time PCR assay for specific detection of novel duck reovirus in China

Zhang

Liu

et al. 2020

BMC Vet Res

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Background: In China, Newly emerging duck reovirus (NDRV) variants have been causing major disease problems in cherry valley ducks. NDRV has the potential to cause high morbidity and 5-50% mortality rates. Severe hemorrhagic-necrosis in the liver and spleen were commonly seen in NDRV affected ducks. The availability of upgraded methods for rapid diagnosis of newly emerging DRV variants is crucial for successful DRV infection control and prevention. Results: In this study, we present a TaqMan-based real-time PCR assay (RT-qPCR) for the detection of NDRV infection. Using the conserved regions within the NDRV genome, we designed the specific primers and probe. The lower limit of detection for NDRV infection was 10 copies/μL (Ct values: 38.3) after the optimization of the RT-qPCR conditions. By cross-checking with other duck viral pathogens, no crossreactivity was observed confirming the assay was highly specific for the detection of NDRV. Reproducibility of the RT-qPCR was confirmed by intra-and inter-assay variability was less than 2.91%(Intra-assay variability of Ct values: 0.07-1.48%; Interassay variability of Ct values: 0.49-2.91%). This RT-qPCR and conventional PCR (cPCR) detected one hundred and twenty samples of NDRV infection from different regions. The result shows that the positive rates were 94.17 and 84.17% respectively. The detection rate of RT-qPCR rapid detection assay was 10% higher than that of the cPCR method. Conclusion: This research developed a highly sensitive, specific, reproducible and versatile of RT-qPCR for quantitatively detecting NDRV. It can be used to study the pathogenesis and epidemiology investigation of NDRV.

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Section: Discussionmentioning

confidence: 99%

A TaqMan-based real-time PCR assay for specific detection of novel duck reovirus in China

Zhang

Liu

et al. 2020

BMC Vet Res

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“…Changes in the phosphorylation levels of the two disease-related signaling pathways might be required for the responses to the C/NDRV infections. Our previous works indicated a central role of the network of seine proteases, consisting of the complement, coagulation and fibrinolytic systems, in the innate immune reactions 28,29 . However, the roles of protein phosphorylation in the responses of serine protease system to the C/NDRV infections are largely unknown.…”

Section: Discussionmentioning

confidence: 99%

“…Minimum number of occurrences was set to 20. Emulate original motif-x was ticked, and other parameters with default 34 28 .…”

Section: Lc-ms/ms Analysismentioning

confidence: 99%

The phosphoproteomic responses of duck (Cairna moschata) to classical/novel duck reovirus infections in the spleen tissue

Yun

Hua

et al. 2020

Sci Rep

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Duck reovirus (DRV) is a fatal member of the genus Orthoreovirus in the family Reoviridae. The disease caused by DRV leads to huge economic losses to the duck industry. Post-translational modification is an efficient strategy to enhance the immune responses to virus infection. However, the roles of protein phosphorylation in the responses of ducklings to Classic/Novel DRV (C/NDRV) infections are largely unknown. Using a high-resolution LC–MS/MS integrated to highly sensitive immune-affinity antibody method, phosphoproteomes of Cairna moschata spleen tissues under the C/NDRV infections were analyzed, producing a total of 8,504 phosphorylation sites on 2,853 proteins. After normalization with proteomic data, 392 sites on 288 proteins and 484 sites on 342 proteins were significantly changed under the C/NDRV infections, respectively. To characterize the differentially phosphorylated proteins (DPPs), a systematic bioinformatics analyses including Gene Ontology annotation, domain annotation, subcellular localization, and Kyoto Encyclopedia of Genes and Genomes pathway annotation were performed. Two important serine protease system-related proteins, coagulation factor X and fibrinogen α-chain, were identified as phosphorylated proteins, suggesting an involvement of blood coagulation under the C/NDRV infections. Furthermore, 16 proteins involving the intracellular signaling pathways of pattern-recognition receptors were identified as phosphorylated proteins. Changes in the phosphorylation levels of MyD88, NF-κB, RIP1, MDA5 and IRF7 suggested a crucial role of protein phosphorylation in host immune responses of C. moschata. Our study provides new insights into the responses of ducklings to the C/NDRV infections at PTM level.

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“…Protein extraction and trypsin digestion were performed as previously described. [ 52 ] Samples were ground in 5 mL tubes and treated using a high intensity ultrasonic processor. Solid material recovered by centrifugation at 4 °C was discarded.…”

Section: Methodsmentioning

confidence: 99%

“…Reconstitution in 0.5 m triethylammonium bicarbonate buffer was done using a 6‐plex TMT Kit (Thermo Fisher Scientific, Waltham, MA, USA) for TMT labeling as previously described. [ 52 ] One unit of TMT reagent was used to label 100 µg of the target sample. The sample‐reagent mixture was left at room temperature for 2 h. Salt was removed and the mixture was dried.…”

Section: Methodsmentioning

confidence: 99%

Comparative Proteomic Analysis Reveals Complex Responses to Bordetella bronchiseptica Infections in the Spleen of Rabbits

et al. 2020

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Bordetella bronchiseptica (B. bronchiseptica) causes a respiratory disease in rabbits. To determine the proteins of B. bronchiseptica in rabbits related to the disease, differentially accumulated proteins in B. bronchiseptica‐infected cells are identified by comparative proteomic analysis. Comparative proteomic analysis detects 5814 proteins and quantifies 4854 of these. Fifty eight upregulated and 38 downregulated proteins are identified in spleen tissue after B. bronchiseptica infection of rabbits (both p < 0.05). The significantly enriched Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways are ribosome, biosynthesis of amino acids, biosynthesis of amino acids, protein export, and carbon metabolism etc. (all p < 0.01). Significantly enriched KEGG pathways include ‘ocu03010 ribosome’ (a); ‘ocu00260 glycine, serine threonine metabolism’. Analyses of control and infected spleen cells detect responses to B. bronchiseptica infection. Many differentially affected proteins are evident, and reflect different biological changes and diverse subcellular localizations between control and infected spleen cells. Infection markedly alters the expressions of proteins linked to the serine protease system, with the ‘phagosome,’ ‘biosynthesis of amino acids,’ ‘glycine, serine threonine metabolism,’ ‘intestinal immune network for IgA production’, and ‘amino sugar and nucleotide sugar metabolism’ associated with B. bronchiseptica infection. The result will inform studies of responses to B. bronchiseptica infections in rabbits.

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Comparative proteomic analysis revealed complex responses to classical/novel duck reovirus infections in Cairna moschata

Cited by 13 publications

References 37 publications

A TaqMan-based real-time PCR assay for specific detection of novel duck reovirus in China

A TaqMan-based real-time PCR assay for specific detection of novel duck reovirus in China

The phosphoproteomic responses of duck (Cairna moschata) to classical/novel duck reovirus infections in the spleen tissue

Comparative Proteomic Analysis Reveals Complex Responses to Bordetella bronchiseptica Infections in the Spleen of Rabbits

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