7 results on '"Chen, Shilong"'
Search Results
2. Development of a duplex SYBR Green I-based quantitative real-time PCR assay for the rapid differentiation of goose and Muscovy duck parvoviruses
- Author
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Fusong Yu, Xiao Shifeng, Xiuqin Chen, Chen Shaoying, Cheng Xiaoxia, Shao Wang, Chen Shilong, and Lin Su
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0301 basic medicine ,GPV ,viruses ,animal diseases ,Oropharynx ,law.invention ,Parvovirus ,chemistry.chemical_compound ,0302 clinical medicine ,Cloaca ,law ,Geese ,Transition Temperature ,Organic Chemicals ,Polymerase chain reaction ,Phylogeny ,biology ,virus diseases ,Viral Load ,Infectious Diseases ,Quantitative Real Time PCR ,Ducks ,Quinolines ,030211 gastroenterology & hepatology ,Muscovy duck parvovirus ,Short Report ,Duplex real-time PCR ,Genome, Viral ,Diamines ,Real-Time Polymerase Chain Reaction ,lcsh:Infectious and parasitic diseases ,MDPV ,Parvoviridae Infections ,03 medical and health sciences ,Goose ,Virology ,biology.animal ,Animals ,lcsh:RC109-216 ,Benzothiazoles ,Poultry Diseases ,Goose parvovirus ,SYBR Green I ,biology.organism_classification ,030104 developmental biology ,chemistry ,Duplex (building) - Abstract
Background Waterfowl parvoviruses, including goose parvovirus (GPV) and Muscovy duck parvovirus (MDPV), can cause seriously diseases in geese and ducks. Developing a fast and precise diagnosis assay for these two parvoviruses is particularly important. Results A duplex SYBR Green I-based quantitative real-time PCR assay was developed for the simultaneous detection and differentiation of GPV and MDPV. The assay yielded melting curves with specific single peak (Tm = 87.3 ± 0.26 °C or Tm = 85.4 ± 0.23 °C) when GPV or MDPV was evaluated, respectively. When both parvoviruses were assessed in one reaction, melting curves with specific double peaks were yielded. Conclusion This duplex quantitative RT-PCR can be used to rapid identify of GPV and MDPV in field cases and artificial trials, which make it a powerful tool for diagnosing, preventing and controlling waterfowl parvovirus infections.
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- 2019
3. Identification of a novel goose parvovirus (GPV) recombinant associated with short beak and dwarfism syndrome in Mainland China, 2015
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Chen Shilong, Lin Fengqiang, You-quang Cheng, Xiao Shifeng, Cheng Xiaoxia, Shao Wang, Nan-yang Wu, Jinxiang Wang, Fusong Yu, Chen Shaoying, and Zhu Xiaoli
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0301 basic medicine ,Microbiology (medical) ,Mainland China ,China ,Dwarfism ,Biology ,Microbiology ,law.invention ,Parvoviridae Infections ,03 medical and health sciences ,Parvovirinae ,law ,Genetics ,medicine ,Animals ,Molecular Biology ,Phylogeny ,Ecology, Evolution, Behavior and Systematics ,Goose parvovirus ,Beak ,medicine.disease ,Virology ,Ducks ,030104 developmental biology ,Infectious Diseases ,Recombinant DNA - Published
- 2016
4. Duckling short beak and dwarfism syndrome virus infection activates host innate immune response involving both DNA and RNA sensors.
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Chen, Shilong, Fang, Tiehui, Xiao, Shifeng, Lin, Fengqiang, Cheng, Xiaoxia, Wang, Shao, Zhu, Xiaoli, Chen, Xiuqin, Zheng, Min, Munir, Muhammad, Huang, Meiqing, Yu, Fusong, and Chen, Shaoying
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VIRUS diseases , *IMMUNE response , *DNA replication , *DUCKLINGS , *DWARFISM - Abstract
Duckling short beak and dwarfism syndrome virus (SBDSV), a newly identified goose parvovirus, causes devastating disease in domestic waterfowl and considerable economic losses to Chinese waterfowl industry. The molecular pathogenesis of SBDSV infection, nature and dynamics of host immune responses against SBDSV infection remained elusive. In this study, we systematically explored the relative mRNA expression profiles of major innate immune-related genes in SBDSV infected duck embryo fibroblasts. We found that SBDSV infection effectively activated host innate immune responses and resulted in significant up-regulation of IFN-β and several vital IFN-stimulated genes (ISGs). These up-regulation responses were mainly attributed to viral genomic DNA and dsRNA replication intermediates. Importantly, the expression of cGAS was significantly induced, whereas the expression of other DNA receptors including DDX41, STING, ZBP1, LSM14A and LRRFIP1 have no significant change. Furthermore, SBDSV infection also activates the up-regulation of TLR3 and inhibited the expression of TLR2 and TLR4; however, no effect was observed on the expression of TLR1, TLR5, TLR7, TLR15 and TLR21. Intriguingly, SBDSV infection significantly up-regulated the expression of RNA sensors such as MDA5 and LGP2, and resulted in a delayed but significant up-regulation of RIG-I gene. Taken together, these data indicate that host multiple sensors including DNA sensor (cGAS) and RNA sensors (TLR3, MDA5 and LGP2) are involved in recognizing a variety of different pathogen associated molecular patterns (PAMPs) including viral genomic ssDNA and dsRNA replication intermediates, which trigger an effective antiviral innate immune response. • SBDSV, a new number of goose parvovirus, causes devastating disease in domestic waterfowl. • The relative mRNA expression profiles of innate immune-related genes in SBDSV infected DEFs were systematically explored. • SBDSV infection activates host innate immune responses and results in significant up-regulation of IFN-β and vital ISGs. • Host multiple sensors are involved in recognizing pathogen associated molecular patterns (PAMPs). [ABSTRACT FROM AUTHOR]
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- 2020
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5. Development and application of a multiplex PCR method for the simultaneous detection of goose parvovirus, waterfowl reovirus, and goose astrovirus in Muscovy ducks.
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Zhang, Shizhong, Dong, Hui, Lin, Fengqiang, Cheng, Xiaoxia, Zhu, Xiaoli, Jiang, Dandan, Xiao, Shifeng, Chen, Shaoying, Chen, Shilong, and Wang, Shao
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GEESE , *WATERFOWL , *DUCKS , *MIXED infections , *REVERSE transcriptase polymerase chain reaction , *POLYMERASE chain reaction - Abstract
A multiplex polymerase chain reaction (PCR) method was developed to detect and distinguish goose parvovirus (GPV), waterfowl reovirus (WRV), and goose astrovirus (GAstV). Three pairs of primers were designed based on conserved regions in the genomic sequences of these enteric viruses and were used to specifically amplify targeted fragments of 493 bp from the viral protein 3 (VP3) gene of GPV, 300 bp from the sigma A-encoding gene of WRV, and 156 bp from the capsid protein-encoding gene of GAstV. The results showed that the primers can specifically amplify target fragments, without any cross-amplification with other viruses, indicating that the method had good specificity. A sensitivity test showed that the detection limit of the multiplex PCR method was 1 × 103 viral copies. A total of 102 field samples from Muscovy ducks with clinically suspected diseases were evaluated using the newly developed multiplex PCR method. The ratio of positive samples to total samples for GPV, WRV, and GAstV was 73.53% (75/102) for multiplex PCR and was 73.53% (75/102) for routine PCR. Seventy-five positive samples were detected by both methods, for a coincidence ratio of 100%. This multiplex PCR method can simultaneously detect GPV, WRV, and GAstV, which are associated with viral enteritis, thereby providing a specific, sensitive, efficient, and accurate new tool for clinical diagnosis and laboratory epidemiological investigations. • A multiplex PCR assay was developed for simultaneous detection for GPV, WRV, and GAstV in Muscovy duck flocks, with good specificity, good accuracy, and high sensitivity. • The multiplex PCR was able to detect at least 1 × 103 viral copies of GPV, WRV, and GAstV. • The multiplex PCR developed is a useful tool for clinical diagnosis of single virus and/or mixed infections in Muscovy duck. [ABSTRACT FROM AUTHOR]
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- 2024
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6. Construction and rescue of Muscovy duck-origin goose parvovirus from an infectious clone containing an E-box deletion within the left terminal region.
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Wang, Shao, Xiao, Shifeng, Cheng, Xiaoxia, Chen, Shaoying, Zhu, Xiaoli, Lin, Fengqiang, and Chen, Shilong
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MUSCOVY duck , *PARVOVIRUS diseases , *EMBRYOS , *CLONING , *POLYMERASE chain reaction - Abstract
Abstract To obtain a deletion mutant of Muscovy duck-origin goose parvovirus (MDGPV) and to analyze its biological characteristics, the pMDGPVPT plasmid, which contains a full-length DNA infectious clone of the MDGPV PT strain, was used in this study as the template. The E-box at nt 315 of the left inverted terminal repeat sequence (L-ITR) was deleted by overlap extension PCR to obtain the infectious recombinant plasmid p-PTΔE315. The p-PTΔE315 plasmid was transfected into 9-day-old non-immune Muscovy duck embryos via the yolk sac and the rescued deletion mutant virus r-PTΔE315 was generated. Experiments to demonstrate the novel deletion mutant virus' biological characteristics showed that r-PTΔE315 can cause typical lesions after infection of Muscovy duck embryos. Compared with its parent strain PT, the virulence of r-PTΔE315 and its proliferation ability in Muscovy duck embryos were attenuated, but its ability to replicate in MDEF cells was enhanced. This study laid the foundation for further understanding of the relationship between E-box deletion in the L-ITR and MDGPV virulence. Highlights • In this study, we constructed a full-length infectious clone (pMDGPVPT). • Deletion of E-box (E315) may improve the adaptability of MDGPV to MDEF cells. • Deletion of E-box (E315) decreased MDGPV replication in Muscovy duck embryos. [ABSTRACT FROM AUTHOR]
- Published
- 2018
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7. Recovery of Muscovy duck-origin goose parvovirus from an infectious clone containing an E-box motif (CACATG) deletion within the left terminal region.
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Wang, Shao, Xiao, Shifeng, Cheng, Xiaoxia, Chen, Shaoying, Zhu, Xiaoli, Lin, Fengqiang, and Chen, Shilong
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PARVOVIRUSES , *PARVOVIRUS B19 , *GEESE , *YOLK sac , *MESENCHYMAL stem cells , *CELL cycle , *VIRAL replication - Abstract
Muscovy duck-origin goose parvovirus (MDGPV) is a causative agent of MDGPV-associated Derzsy's disease. To evalute the role of the cis -acting element E-box (CACATG) deletion on MDGPV eplication, an infectious plasmid clone p-PTΔE287, having one E-box deletion at nucleotide (nt) 287 of the left inverted terminal repeat sequence (L-ITR), was constructed by overlap extension PCR deleting the 287CACATG292 motif from the plasmid pMDGPVPT containing the full-length genome of the virulent MDGPV strain PT. The p-PTΔE287 plasmid was transfected into 9-day-old non-immune Muscovy duck embryos via the yolk sac, resulting in successful rescue of the deletion mutant virus r-PTΔE287. Compared with its parental virus PT, the virulence and the replication ability of r-PTΔE287 were reduced. In addition, we examined the ability of r-PTΔE287 to manipulate cell cycle progression. The results showed that r-PTΔE287 replication results in G0/G1 phase accumulation of infected duck embryo liver mesenchymal stem cells (BMSCs) and that this accumulation is caused by the prevention of cell cycle entry from G0/G1 phase into S phase. Taken together, introducing 287CACATG292 element deletion into MDGPV PT genomic DNA that induced rescued mutant virus (r-PTΔE287) cell cycle arrest function at the G0/G1 phase, which might inhibit MDGPV replication and virus progeny production. This study laid the foundation for further understanding of the relationship between E-box deletion in the L-ITR and MDGPV virulence. • We constructed an E-box motif (CACATG)-deleted MDGPV. • Deletion of E-box (E287) decreased MDGPV replication in Muscovy duck embryos. • The mutant virus r-PTΔE287 may induce cell cycle arrest at G0/G1 phase. [ABSTRACT FROM AUTHOR]
- Published
- 2019
- Full Text
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