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Article ; Online: Establishment and Application of a Triplex Real-Time RT-PCR Assay for Differentiation of PEDV, PoRV, and PDCoV.

Hou, Wenwen / Fan, Maodi / Zhu, Zhenbang / Li, Xiangdong

2023 Volume 15, Issue 6

Abstract: Porcine viral diarrhea is very common in clinical practice and has caused huge losses to the pig industry. Porcine epidemic diarrhea virus (PEDV), porcine rotavirus (PoRV), and porcine deltacoronavirus (PDCoV) are important pathogens of porcine viral ... ...

Abstract	Porcine viral diarrhea is very common in clinical practice and has caused huge losses to the pig industry. Porcine epidemic diarrhea virus (PEDV), porcine rotavirus (PoRV), and porcine deltacoronavirus (PDCoV) are important pathogens of porcine viral diarrhea. Co-infection situations among these three viruses in clinics are common, which increases the difficulty of differential diagnosis. Currently, polymerase chain reaction (PCR) is commonly used to detect pathogens. TaqMan real-time PCR is more sensitive than conventional PCR and has better specificity and accuracy. In this study, a triplex real-time RT-PCR assay based on TaqMan probes was developed for differential detection of PEDV, PoRV, and PDCoV. The triplex real-time RT-PCR assay developed in this study could not detect unrelated pathogens and showed satisfactory specificity, sensitivity, repeatability, and reproducibility with a limit of detection (LOD) of 6.0 × 10
MeSH term(s)	Animals ; Swine ; Porcine epidemic diarrhea virus/genetics ; Real-Time Polymerase Chain Reaction/veterinary ; Real-Time Polymerase Chain Reaction/methods ; Coronavirus Infections/diagnosis ; Coronavirus Infections/veterinary ; Coronavirus Infections/epidemiology ; Reverse Transcriptase Polymerase Chain Reaction ; Coinfection/diagnosis ; Coinfection/veterinary ; Reproducibility of Results ; Swine Diseases ; Diarrhea/diagnosis ; Diarrhea/veterinary ; Sensitivity and Specificity
Language	English
Publishing date	2023-05-25
Publishing country	Switzerland
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2516098-9
ISSN	1999-4915 ; 1999-4915
ISSN (online)	1999-4915
ISSN	1999-4915
DOI	10.3390/v15061238
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Tubercidin inhibits PRRSV replication via RIG-I/NF-κB pathways and interrupting viral nsp2 synthesis.

Xu, Yuqian / Zhu, Zhenbang / Zhang, Meng / Chen, Lulu / Tian, Kegong / Li, Xiangdong

Microbiology spectrum

2024 Volume 12, Issue 3, Page(s) e0347923

Abstract: Porcine reproductive and respiratory syndrome virus (PRRSV) is an RNA virus with constantly emerging recombinant and mutant strains. Because of the high genetic diversity of PRRSV, current vaccines only provide partial protection against the infection of ...

Abstract	Porcine reproductive and respiratory syndrome virus (PRRSV) is an RNA virus with constantly emerging recombinant and mutant strains. Because of the high genetic diversity of PRRSV, current vaccines only provide partial protection against the infection of heterologous strains, which makes it a challenge for PRRSV prevention and control. Tubercidin is a naturally extracted compound with potential antiviral properties. However, whether tubercidin has anti-PRRSV ability is unknown. Our study found that tubercidin showed effective antiviral effects on PRRSV replication. In terms of mechanism, tubercidin suppressed PRRSV at the entry, replication, and release steps of the viral life cycle. Additionally, we demonstrated that tubercidin treatment promoted the activation of retinoic acid-inducible gene I and nuclear factor kappa-light-chain-enhancer of activated B cell signaling pathway, thus increasing the type I interferon and inflammatory cytokine expression. Furthermore, tubercidin restrained the viral non-structural protein 2 expression and viral dsRNA synthesis and ultimately inhibited PRRSV replication. Hence, our data showed that tubercidin is promising and has potential antiviral ability against PRRSV replication Importance: Porcine reproductive and respiratory syndrome (PRRS) is one of the most important swine diseases, which causes huge economic loss worldwide. However, there is no effective therapeutic method for PRRS prevention and control. Here, we found that tubercidin, a naturally extracted adenosine analog, exhibited strong anti-porcine reproductive and respiratory syndrome virus (PRRSV) activity. Mechanically, tubercidin inhibited viral binding, replication, and release. Tubercidin suppressed PRRSV non-structural protein 2 expression, which is important for the formation of replication and transcription complex, leading to the block of viral RNA synthesis and PRRSV replication. Moreover, tubercidin could activate retinoic acid-inducible gene I/nuclear factor kappa-light-chain-enhancer of activated B cell innate immune signaling pathway and increased the expression of interferons and proinflammatory cytokines, which was the other way to inhibit PRRSV replication. Our work evaluated the potential value of tubercidin as an antiviral agent on PRRSV replication and provided a new way to prevent PRRSV replication
MeSH term(s)	Swine ; Animals ; Porcine respiratory and reproductive syndrome virus/metabolism ; NF-kappa B/metabolism ; Porcine Reproductive and Respiratory Syndrome ; Tubercidin ; Cytokines/metabolism ; DEAD Box Protein 58 ; Antiviral Agents/pharmacology ; Tretinoin
Chemical Substances	NF-kappa B ; Tubercidin (M351LCX45Y) ; Cytokines ; DEAD Box Protein 58 (EC 3.6.4.13) ; Antiviral Agents ; Tretinoin (5688UTC01R)
Language	English
Publishing date	2024-02-01
Publishing country	United States
Document type	Journal Article
ZDB-ID	2807133-5
ISSN	2165-0497 ; 2165-0497
ISSN (online)	2165-0497
ISSN	2165-0497
DOI	10.1128/spectrum.03479-23
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Antagonisms of ASFV towards Host Defense Mechanisms: Knowledge Gaps in Viral Immune Evasion and Pathogenesis.

Yu, Liangzheng / Zhu, Zhenbang / Deng, Junhua / Tian, Kegong / Li, Xiangdong

Viruses

2023 Volume 15, Issue 2

Abstract: African swine fever (ASF) causes high morbidity and mortality of both domestic pigs and wild boars and severely impacts the swine industry worldwide. ASF virus (ASFV), the etiologic agent of ASF epidemics, mainly infects myeloid cells in swine ... ...

Abstract	African swine fever (ASF) causes high morbidity and mortality of both domestic pigs and wild boars and severely impacts the swine industry worldwide. ASF virus (ASFV), the etiologic agent of ASF epidemics, mainly infects myeloid cells in swine mononuclear phagocyte system (MPS), including blood-circulating monocytes, tissue-resident macrophages, and dendritic cells (DCs). Since their significant roles in bridging host innate and adaptive immunity, these cells provide ASFV with favorable targets to manipulate and block their antiviral activities, leading to immune escape and immunosuppression. To date, vaccines are still being regarded as the most promising measure to prevent and control ASF outbreaks. However, ASF vaccine development is delayed and limited by existing knowledge gaps in viral immune evasion, pathogenesis, etc. Recent studies have revealed that ASFV can employ diverse strategies to interrupt the host defense mechanisms via abundant self-encoded proteins. Thus, this review mainly focuses on the antagonisms of ASFV-encoded proteins towards IFN-I production, IFN-induced antiviral response, NLRP3 inflammasome activation, and GSDMD-mediated pyroptosis. Additionally, we also make a brief discussion concerning the potential challenges in future development of ASF vaccine.
MeSH term(s)	Animals ; Swine ; African Swine Fever ; Immune Evasion ; Monocytes ; Adaptive Immunity ; Antiviral Agents ; Sus scrofa
Chemical Substances	Antiviral Agents
Language	English
Publishing date	2023-02-19
Publishing country	Switzerland
Document type	Journal Article ; Review ; Research Support, Non-U.S. Gov't
ZDB-ID	2516098-9
ISSN	1999-4915 ; 1999-4915
ISSN (online)	1999-4915
ISSN	1999-4915
DOI	10.3390/v15020574
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Bergamottin Inhibits PRRSV Replication by Blocking Viral Non-Structural Proteins Expression and Viral RNA Synthesis.

Zhu, Zhenbang / Xu, Yuqian / Chen, Lulu / Zhang, Meng / Li, Xiangdong

Viruses

2023 Volume 15, Issue 6

Abstract: The porcine reproductive and respiratory syndrome virus (PRRSV) causes economic losses in the swine industry worldwide. However, current vaccines cannot provide effective protection against PRRSV, and PRRSV-specific treatments for infected herds are ... ...

Abstract	The porcine reproductive and respiratory syndrome virus (PRRSV) causes economic losses in the swine industry worldwide. However, current vaccines cannot provide effective protection against PRRSV, and PRRSV-specific treatments for infected herds are still unavailable. In this study, we found that bergamottin showed strong inhibitory effects against PRRSV replication. Bergamottin inhibited PRRSV at the stage of the replication cycle. Mechanically, bergamottin promoted the activation of IRF3 and NF-κB signaling, leading to the increased expression of proinflammatory cytokines and interferon, which inhibited viral replication to some extent. In addition, bergamottion could reduce the expression of the non-structural proteins (Nsps), leading to the interruption of replication and transcription complex (RTC) formation and viral dsRNA synthesis, ultimately restraining PRRSV replication. Our study identified that bergamottin possesses potential value as an antiviral agent against PRRSV in vitro.
MeSH term(s)	Animals ; Swine ; Porcine respiratory and reproductive syndrome virus/metabolism ; Cell Line ; Furocoumarins/pharmacology ; Viral Nonstructural Proteins/metabolism ; Virus Replication ; Porcine Reproductive and Respiratory Syndrome
Chemical Substances	bergamottin (JMU611YFRB) ; Furocoumarins ; Viral Nonstructural Proteins
Language	English
Publishing date	2023-06-13
Publishing country	Switzerland
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2516098-9
ISSN	1999-4915 ; 1999-4915
ISSN (online)	1999-4915
ISSN	1999-4915
DOI	10.3390/v15061367
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article: Development and application of a duplex real‐time PCR assay for differentiation of genotypes I and II African swine fever viruses

Li, Xiangdong / Hu, Yongxin / Liu, Penggang / Zhu, Zhenbang / Liu, Paorao / Chen, Changhai / Wu, Xiaodong

Transboundary and emerging diseases. 2022 Sept., v. 69, no. 5

2022

Abstract: Genotype II African swine fever virus (ASFV) has been plaguing Chinese pig industry and caused severe morbidity and mortality of pigs resulting in huge economic losses since its first report in August 2018. Most recently, two genotype I ASFVs with low ... ...

Abstract	Genotype II African swine fever virus (ASFV) has been plaguing Chinese pig industry and caused severe morbidity and mortality of pigs resulting in huge economic losses since its first report in August 2018. Most recently, two genotype I ASFVs with low virulence but efficient transmissibility in pigs were reported in China, which makes the diagnosis and control of this lethal disease more challenging. Therefore, it is prerequisite and important to differentiate genotype I from genotype II upon ASFV outbreaks before making any stringent control procedures. In this study, a duplex real‐time PCR assay based on ASFV E296R gene was established which could simultaneously detect genotypes I and II ASFVs with two pairs of primers and two probes. Plasmid containing ASFV genes was used to test the sensitivity, repeatability, and reproducibility. DNA or cDNA samples of ASFV and other swine viruses were used to test the specificity. The results showed that the established duplex real‐time PCR assay has satisfied specificity, sensitivity, repeatability, and reproducibility. In addition, the assay was applied to differentiate 84 ASFV positive clinical samples including lymph nodes, spleen, kidney, lung, liver, blood, nasal swab, and environmental swab samples which were sent to National ASF Reference Laboratory from April 2020 to September 2021. The results showed that all these ASFV positive samples belong to genotype II ASFV. The established duplex real‐time PCR in this study provides a powerful tool for rapid detection and differentiation between genotypes I and II ASFVs and will facilitate efficient control of ASFV in China.
Keywords	African swine fever virus ; blood ; genes ; genotype ; industry ; kidneys ; liver ; lungs ; lymph ; morbidity ; mortality ; nose ; plasmids ; quantitative polymerase chain reaction ; rapid methods ; spleen ; stress response ; swine ; virulence ; China
Language	English
Dates of publication	2022-09
Size	p. 2971-2979.
Publishing place	John Wiley & Sons, Ltd
Document type	Article
Note	JOURNAL ARTICLE
ZDB-ID	2414822-2
ISSN	1865-1682 ; 1865-1674
ISSN (online)	1865-1682
ISSN	1865-1674
DOI	10.1111/tbed.14459
Database	NAL-Catalogue (AGRICOLA)

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Article: Metabolomics Analysis of PK-15 Cells with Pseudorabies Virus Infection Based on UHPLC-QE-MS

Liu, Panrao / Hu, Danhe / Yuan, Lili / Lian, Zhengmin / Yao, Xiaohui / Zhu, Zhenbang / Li, Xiangdong

Viruses. 2022 May 27, v. 14, no. 6

2022

Abstract: Viruses depend on the metabolic mechanisms of the host to support viral replication. We utilize an approach based on ultra-high-performance liquid chromatography/Q Exactive HF-X Hybrid Quadrupole-Orbitrap Mass (UHPLC-QE-MS) to analyze the metabolic ... ...

Abstract	Viruses depend on the metabolic mechanisms of the host to support viral replication. We utilize an approach based on ultra-high-performance liquid chromatography/Q Exactive HF-X Hybrid Quadrupole-Orbitrap Mass (UHPLC-QE-MS) to analyze the metabolic changes in PK-15 cells induced by the infections of the pseudorabies virus (PRV) variant strain and Bartha K61 strain. Infections with PRV markedly changed lots of metabolites, when compared to the uninfected cell group. Additionally, most of the differentially expressed metabolites belonged to glycerophospholipid metabolism, sphingolipid metabolism, purine metabolism, and pyrimidine metabolism. Lipid metabolites account for the highest proportion (around 35%). The results suggest that those alterations may be in favor of virion formation and genome amplification to promote PRV replication. Different PRV strains showed similar results. An understanding of PRV-induced metabolic reprogramming will provide valuable information for further studies on PRV pathogenesis and the development of antiviral therapy strategies.
Keywords	Suid alphaherpesvirus 1 ; gene expression regulation ; genome ; glycerophospholipids ; hybrids ; metabolites ; metabolomics ; pathogenesis ; sphingolipids ; therapeutics ; ultra-performance liquid chromatography ; virion ; virus replication
Language	English
Dates of publication	2022-0527
Publishing place	Multidisciplinary Digital Publishing Institute
Document type	Article
ZDB-ID	2516098-9
ISSN	1999-4915
ISSN	1999-4915
DOI	10.3390/v14061158
Database	NAL-Catalogue (AGRICOLA)

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Article: Isolation and genomic characterization of a Chinese NADC34‐like PRRSV isolated from Jiangsu province

Zhu, Zhenbang / Yuan, Lili / Hu, Danhe / Lian, Zhengmin / Yao, Xiaohui / Liu, Panrao / Li, Xiangdong

Transboundary and emerging diseases. 2022 July, v. 69, no. 4

2022

Abstract: Porcine reproductive and respiratory syndrome virus (PRRSV) is one of the most important causative agents to swine industry, which has been epidemic more than 30 years. The emergence and recombination of new virus strains bring great challenges to the ... ...

Abstract	Porcine reproductive and respiratory syndrome virus (PRRSV) is one of the most important causative agents to swine industry, which has been epidemic more than 30 years. The emergence and recombination of new virus strains bring great challenges to the prevention and control of PRRSV. In the present study, we reported and characterized a novel PRRSV strain, designated as JS2021NADC34, which was for the first time isolated from clinical samples in Jiangsu province, China. Phylogenetic analysis demonstrated that JS2021NADC34 belonging to sublineage 1.5 of PRRSV‐2 and was highly related to NADC34‐like strains. Genetically, JS2021NADC34 strain had a continuous 100 aa depletion in NSP2, as compared to VR‐2332 strain, which was consistent with most reported NADC34‐like strains. Moreover, there were several amino acid substitutions occurred in the antigenic regions of GP2–GP5. Similar to other reported NADC34‐like PRRSV in China, JS2021NADC34 had no recombination with other domestic strains, which indicates this sublineage of PRRSV may be directly transported from the United States and have not undergone extensive mutation and recombination with local strains yet.
Keywords	Betaarterivirus suid 2 ; amino acids ; genomics ; phylogeny ; porcine reproductive and respiratory syndrome ; pork industry ; viruses ; China
Language	English
Dates of publication	2022-07
Size	p. e1015-e1027.
Publishing place	John Wiley & Sons, Ltd
Document type	Article
Note	JOURNAL ARTICLE
ZDB-ID	2414822-2
ISSN	1865-1682 ; 1865-1674
ISSN (online)	1865-1682
ISSN	1865-1674
DOI	10.1111/tbed.14392
Database	NAL-Catalogue (AGRICOLA)

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Article ; Online: Metabolomics Analysis of PK-15 Cells with Pseudorabies Virus Infection Based on UHPLC-QE-MS.

Liu, Panrao / Hu, Danhe / Yuan, Lili / Lian, Zhengmin / Yao, Xiaohui / Zhu, Zhenbang / Li, Xiangdong

Viruses

2022 Volume 14, Issue 6

Abstract	Viruses depend on the metabolic mechanisms of the host to support viral replication. We utilize an approach based on ultra-high-performance liquid chromatography/Q Exactive HF-X Hybrid Quadrupole-Orbitrap Mass (UHPLC-QE-MS) to analyze the metabolic changes in PK-15 cells induced by the infections of the pseudorabies virus (PRV) variant strain and Bartha K61 strain. Infections with PRV markedly changed lots of metabolites, when compared to the uninfected cell group. Additionally, most of the differentially expressed metabolites belonged to glycerophospholipid metabolism, sphingolipid metabolism, purine metabolism, and pyrimidine metabolism. Lipid metabolites account for the highest proportion (around 35%). The results suggest that those alterations may be in favor of virion formation and genome amplification to promote PRV replication. Different PRV strains showed similar results. An understanding of PRV-induced metabolic reprogramming will provide valuable information for further studies on PRV pathogenesis and the development of antiviral therapy strategies.
MeSH term(s)	Animals ; Chromatography, High Pressure Liquid ; Herpesvirus 1, Suid/genetics ; Metabolomics ; Pseudorabies ; Swine ; Swine Diseases
Language	English
Publishing date	2022-05-27
Publishing country	Switzerland
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2516098-9
ISSN	1999-4915 ; 1999-4915
ISSN (online)	1999-4915
ISSN	1999-4915
DOI	10.3390/v14061158
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Development and application of a duplex real-time PCR assay for differentiation of genotypes I and II African swine fever viruses.

Li, Xiangdong / Hu, Yongxin / Liu, Penggang / Zhu, Zhenbang / Liu, Paorao / Chen, Changhai / Wu, Xiaodong

Transboundary and emerging diseases

2022 Volume 69, Issue 5, Page(s) 2971–2979

Abstract	Genotype II African swine fever virus (ASFV) has been plaguing Chinese pig industry and caused severe morbidity and mortality of pigs resulting in huge economic losses since its first report in August 2018. Most recently, two genotype I ASFVs with low virulence but efficient transmissibility in pigs were reported in China, which makes the diagnosis and control of this lethal disease more challenging. Therefore, it is prerequisite and important to differentiate genotype I from genotype II upon ASFV outbreaks before making any stringent control procedures. In this study, a duplex real-time PCR assay based on ASFV E296R gene was established which could simultaneously detect genotypes I and II ASFVs with two pairs of primers and two probes. Plasmid containing ASFV genes was used to test the sensitivity, repeatability, and reproducibility. DNA or cDNA samples of ASFV and other swine viruses were used to test the specificity. The results showed that the established duplex real-time PCR assay has satisfied specificity, sensitivity, repeatability, and reproducibility. In addition, the assay was applied to differentiate 84 ASFV positive clinical samples including lymph nodes, spleen, kidney, lung, liver, blood, nasal swab, and environmental swab samples which were sent to National ASF Reference Laboratory from April 2020 to September 2021. The results showed that all these ASFV positive samples belong to genotype II ASFV. The established duplex real-time PCR in this study provides a powerful tool for rapid detection and differentiation between genotypes I and II ASFVs and will facilitate efficient control of ASFV in China.
MeSH term(s)	African Swine Fever/diagnosis ; African Swine Fever/epidemiology ; African Swine Fever Virus/genetics ; Animals ; DNA, Complementary ; DNA, Viral/genetics ; Genotype ; Real-Time Polymerase Chain Reaction/methods ; Real-Time Polymerase Chain Reaction/veterinary ; Reproducibility of Results ; Swine ; Swine Diseases
Chemical Substances	DNA, Complementary ; DNA, Viral
Language	English
Publishing date	2022-01-28
Publishing country	Germany
Document type	Journal Article
ZDB-ID	2414822-2
ISSN	1865-1682 ; 1865-1674
ISSN (online)	1865-1682
ISSN	1865-1674
DOI	10.1111/tbed.14459
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: LGP2 Promotes Type I Interferon Production To Inhibit PRRSV Infection via Enhancing MDA5-Mediated Signaling.

Zhu, Zhenbang / Zhang, Meng / Yuan, Lili / Xu, Yuqian / Zhou, Han / Lian, Zhengmin / Liu, Panrao / Li, Xiangdong

Journal of virology

2023 Volume 97, Issue 1, Page(s) e0184322

Abstract: Porcine reproductive and respiratory syndrome virus (PRRSV) is one of the most important pathogens in the global pig industry, which modulates the host's innate antiviral immunity to achieve immune evasion. RIG-I-like receptors (RLRs) sense viral RNA and ...

Abstract	Porcine reproductive and respiratory syndrome virus (PRRSV) is one of the most important pathogens in the global pig industry, which modulates the host's innate antiviral immunity to achieve immune evasion. RIG-I-like receptors (RLRs) sense viral RNA and activate the interferon signaling pathway. LGP2, a member of the RLR family, plays an important role in regulating innate immunity. However, the role of LGP2 in virus infection is controversial. Whether LGP2 has a role during infection with PRRSV remains unclear. Here, we found that LGP2 overexpression restrained the replication of PRRSV, while LGP2 silencing facilitated PRRSV replication. LGP2 was prone to interact with MDA5 and enhanced viral RNA enrichment and recognition by MDA5, thus promoting the activation of RIG-I/IRF3 and NF-κB signaling pathways and reinforcing the expression of proinflammatory cytokines and type I interferon during PRRSV infection. Meanwhile, there was a decreased protein expression of LGP2 upon PRRSV infection
MeSH term(s)	Animals ; Immunity, Innate ; Interferon Type I ; NF-kappa B/metabolism ; Porcine respiratory and reproductive syndrome virus/genetics ; RNA Helicases/metabolism ; RNA, Viral/genetics ; Signal Transduction/genetics ; Swine ; Porcine Reproductive and Respiratory Syndrome/immunology
Chemical Substances	Interferon Type I ; NF-kappa B ; RNA Helicases (EC 3.6.4.13) ; RNA, Viral
Language	English
Publishing date	2023-01-09
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	80174-4
ISSN	1098-5514 ; 0022-538X
ISSN (online)	1098-5514
ISSN	0022-538X
DOI	10.1128/jvi.01843-22
Database	MEDical Literature Analysis and Retrieval System OnLINE

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