An efficient peptide-based ELISA for differentiating fowl adenovirus 4–infected chickens from vaccinated chickens

2021 ◽  
pp. 104063872110057
Author(s):  
Songhua Xie ◽  
Qiuping Shen ◽  
Wei Zhang ◽  
Weikang Wang ◽  
Quan Xie ◽  
...  
Keyword(s):  
Indirect Elisa ◽  
Nonstructural Protein ◽  
Immunofluorescence Assay ◽  
Economic Losses ◽  
Clinical Samples ◽  
Immunogenic Peptide ◽  
Hydropericardium Syndrome ◽  
Fowl Adenovirus ◽  
Inactivated Vaccines ◽  
Fowl Adenovirus Serotype 4

Fowl adenovirus serotype 4 (FAdV4), the causative agent of hepatitis-hydropericardium syndrome (HPS), has caused major economic losses to the poultry industry worldwide. Although inactivated vaccines have been deployed widely against FAdV4, a DIVA (differentiating infected from vaccinated animals) test specific for FAdV4 has not been available. We synthesized an immunogenic peptide, corresponding to regions 66–88 aa of the 22K nonstructural protein of FAdV4, and used the peptide as coating antigen to develop an indirect ELISA for a DIVA test specific to FAdV4. Specificity analysis showed that the ELISA only reacted with sera against FAdV4, and not with sera against other pathogens tested. Moreover, the ELISA could effectively differentiate FAdV4–infected chickens from vaccinated chickens. In a test of sera from experimentally infected chickens, the ELISA had 95% and 85% concordance with an indirect immunofluorescence assay (indirect IFA) and a commercial ELISA, respectively, and the concordance was 80.5% between the ELISA and the indirect IFA in detecting clinical infection samples. Our peptide-based ELISA provides an efficient DIVA test for FAdV4 in clinical samples.

scholarly journals Requirement of Cellular Protein CCT7 for the Replication of Fowl Adenovirus Serotype 4 (FAdV-4) in Leghorn Male Hepatocellular Cells Via Interaction with the Viral Hexon Protein

Viruses ◽  
2019 ◽  
Vol 11 (2) ◽  
pp. 107 ◽  
Author(s):  
Junfeng Gao ◽  
Mingliang Zhao ◽  
Xueyan Duan ◽  
Yongqiang Wang ◽  
Hong Cao ◽  
...  
Keyword(s):  
Molecular Mechanisms ◽  
Ectopic Expression ◽  
Cellular Protein ◽  
Economic Losses ◽  
Adenovirus Serotype ◽  
Hydropericardium Syndrome ◽  
Fowl Adenovirus ◽  
Infected Cells ◽  
Lmh Cells ◽  
Fowl Adenovirus Serotype 4

Fowl adenovirus serotype 4 (FAdV-4) causes hepatitis-hydropericardium syndrome (HHS), leading to severe economic losses in the poultry industry. Although the pathogenesis of FAdV-4 infection has caused much attention, the underlying molecular mechanisms remain poorly understood. Here, we identified chaperonin containing TCP-1 subunit eta (CCT7) as an interacting partner of the FAdV-4 capsid protein hexon. We found that ectopic expression of CCT7 in leghorn male hepatocellular (LMH) cells enhanced hexon expression in pRK5-flag-hexon transfected cells. On the contrary, knockdown of cellular CCT7 by RNAi markedly reduced hexon expression in FAdV-4-infected cells and suppressed viral replication. These data suggest that CCT7 is required for FAdV-4 replication and may serve as a potential target for controlling FAdV-4 infection.

scholarly journals Identification of a novel immunological epitope on Hexon of fowl adenovirus serotype 4

AMB Express ◽  
2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Jingyi Liu ◽  
Nan Mei ◽  
Yaling Wang ◽  
Xinjin Shi ◽  
Hongjun Chen
Keyword(s):  
Amino Acid Sequence Analysis ◽  
Serum Samples ◽  
Protein Amino Acid ◽  
Adenovirus Serotype ◽  
Linear Peptide ◽  
Hydropericardium Syndrome ◽  
Hexon Protein ◽  
Fowl Adenovirus ◽  
Fowl Adenovirus Serotype 4

AbstractFowl adenovirus serotype 4 (FAdV-4), the causative agent of hepatitis-hydropericardium syndrome (HHS), distributed widely in the poultry farms in China. Hexon is one of the major capsid proteins associated with viral species or serotypes. However, the epitopes of Hexon protein remain largely unknown. In this study, a monoclonal antibody (mAb) specific to Hexon protein of FAdV-4, designated as 3G8, was generated. Subsequently, the linear peptide recognized by 3G8 was mapped and identified as 213AYGAYVK219 using a series of overlapping peptides generated from Hexon protein. Amino acid sequence analysis revealed that the epitope recognized by 3G8 was highly conserved across all the FAdVs. The epitope was immunogenic and could be recognized by FAdV-4 positive chicken serum samples. These findings will enrich our knowledge regarding the epitope on Hexon and provide valuable information for further characterization of the antigenicity of Hexon protein.

scholarly journals Development of a Novel Avian Vaccine Vector Derived From the Emerging Fowl Adenovirus 4

2021 ◽  
Vol 12 ◽  
Author(s):  
Qing Pan ◽  
Yu Zhang ◽  
Aijing Liu ◽  
Hongyu Cui ◽  
Yulong Gao ◽  
...  
Keyword(s):  
Fluorescent Protein ◽  
Virus Genome ◽  
Open Reading Frames ◽  
Vaccine Vector ◽  
Economic Losses ◽  
Foreign Gene ◽  
The Novel ◽  
Hydropericardium Syndrome ◽  
Fowl Adenovirus ◽  
Green Fluorescent

Severe hepatitis-hydropericardium syndrome (HHS) associated with a novel viral genotype, fowl adenovirus 4 (FAdV-4), has emerged and widely spread in China since 2015, causing severe economic losses to the poultry industry. We previously reported that the hexon gene is responsible for pathogenicity and obtained a non-pathogenic hexon-replacement rHN20 strain; however, the lack of information about the non-essential regions for virus replication limits the development of a FAdV-4 vector. This study first established an enhanced green fluorescent protein (EGFP)-indicator virus based on the FAdV-4 reverse genetic technique, effective for batch operations in the virus genome. Based on this, 10 open reading frames (ORFs) at the left end and 13 ORFs at the right end of the novel FAdV-4 genome were deleted separately and identified as non-essential genes for viral replication, providing preliminary insertion sites for foreign genes. To further improve its feasibility as a vaccine vector, seven combinations of ORFs were successfully replaced with EGFP without affecting the immunogenicity of the vector backbone. Finally, a recombinant rHN20-vvIBDV-VP2 strain, expressing the VP2 protein of very virulent infectious bursa disease virus (vvIBDV), was rescued and showed complete protection against FAdV-4 and vvIBDV. Thus, the novel FAdV-4 vector could provide sufficient protection for HHS and efficient exogenous gene delivery. Overall, our findings systemically identified 23 non-essential ORFs for FAdV-4 replication and seven foreign gene insertion regions, providing valuable information for an in-depth understanding of the novel FAdV-4 pathogenesis and development of multivalent vaccines.

scholarly journals Airborne Transmission of a Serotype 4 Fowl Adenovirus in Chickens

Viruses ◽  
2019 ◽  
Vol 11 (3) ◽  
pp. 262 ◽  
Author(s):  
Gang Li ◽  
Guanliu Yu ◽  
Yujuan Niu ◽  
Yumei Cai ◽  
Sidang Liu
Keyword(s):  
Infectious Disease ◽  
Infected Group ◽  
Economic Losses ◽  
Control Group ◽  
Airborne Transmission ◽  
Hydropericardium Syndrome ◽  
Fowl Adenovirus ◽  
Viral Aerosol ◽  
Subclinical Symptoms ◽  
And Control

Serotype 4 fowl adenovirus (FAdV-4) is the main pathogen for hydropericardium syndrome (HPS) in chickens. It has caused major economic losses in the global poultry industry. Currently, FAdV-4′s transmission routes in chickens remain unclear. Here we investigate the airborne transmission routes of FAdV-4 in chickens. A total of 45 ten-day-old chickens were equally divided into three groups (infected group/isolator A, airborne group/isolator B, and control group/isolator C). Of note, isolators A and B were connected by a leak-free pipe. The results showed that the virus could form a viral aerosol, detected in isolators two days post infection (dpi). The viral aerosol reached a peak at 4 dpi in the infected group. Healthy chickens in the airborne group were infected by the virus at 8 dpi. The chickens of the airborne group demonstrated subclinical symptoms capable of shedding the virus for some time. This finding suggests that FAdV-4 can be efficiently transmitted among chickens by aerosol transmission. These findings have significant implications for developing strategies to control this infectious disease epidemic.

scholarly journals Pathogenesis of Hypervirulent Fowl Adenovirus Serotype 4: The Contributions of Viral and Host Factors

Viruses ◽  
2019 ◽  
Vol 11 (8) ◽  
pp. 741 ◽  
Author(s):  
Zeng Wang ◽  
Jun Zhao
Keyword(s):  
Host Defense ◽  
Host Factors ◽  
Present Review ◽  
Defense System ◽  
Adenovirus Serotype ◽  
Vaccine Candidates ◽  
Hydropericardium Syndrome ◽  
Fowl Adenovirus ◽  
Host Defense System ◽  
Fowl Adenovirus Serotype 4

Since 2015, severe outbreaks of hepatitis-hydropericardium syndrome (HHS), caused by hypervirulent fowl adenovirus serotype 4 (FAdV-4), have emerged in several provinces in China, posing a great threat to poultry industry. So far, factors contributing to the pathogenesis of hypervirulent FAdV-4 have not been fully uncovered. Elucidation of the pathogenesis of FAdV-4 will facilitate the development of effective FAdV-4 vaccine candidates for the control of HHS and vaccine vector. The interaction between pathogen and host defense system determines the pathogenicity of the pathogen. Therefore, the present review highlights the knowledge of both viral and host factors contributing to the pathogenesis of hypervirulent FAdV-4 strains to facilitate the related further studies.

Recombinant Fiber-2 Protein-Based Indirect ELISA for Antibody Detection of Fowl Adenovirus Serotype 4

2018 ◽  
Vol 62 (1) ◽  
pp. 73-78 ◽  
Author(s):  
Zi-rong He ◽  
Si-fan Ruan ◽  
Jing Zhao ◽  
Hui-ming Yang ◽  
Guo-zhong Zhang
Keyword(s):  
Indirect Elisa ◽  
Antibody Detection ◽  
Adenovirus Serotype ◽  
Fowl Adenovirus ◽  
Fowl Adenovirus Serotype 4

scholarly journals Hydropericardium syndrome caused by fowl adenovirus serotype 4 in replacement pullets

2019 ◽  
Vol 81 (2) ◽  
pp. 245-251 ◽  
Author(s):  
Zhen CHEN ◽  
Shaohua SHI ◽  
Baoming QI ◽  
Su LIN ◽  
Cuiteng CHEN ◽  
...  
Keyword(s):  
Adenovirus Serotype ◽  
Hydropericardium Syndrome ◽  
Fowl Adenovirus ◽  
Fowl Adenovirus Serotype 4

scholarly journals Identification, Pathogenicity of Novel Fowl Adenovirus Serotype 4 SDJN0105 in Shandong, China and Immunoprotective Evaluation of the Newly Developed Inactivated Oil-emulsion FAdV-4 Vaccine

Viruses ◽  
2019 ◽  
Vol 11 (7) ◽  
pp. 627 ◽  
Author(s):  
Kai Meng ◽  
Xiaoyuan Yuan ◽  
Jiang Yu ◽  
Yuxia Zhang ◽  
Wu Ai ◽  
...  
Keyword(s):  
Clinical Symptoms ◽  
Histopathological Examination ◽  
Lethal Dose ◽  
Economic Losses ◽  
Effective Vaccine ◽  
Drug Candidate ◽  
Adenovirus Serotype ◽  
Oil Emulsion ◽  
Fowl Adenovirus ◽  
Fowl Adenovirus Serotype 4

Since mid-2015, numerous outbreaks of hydropericardium-hepatitis syndrome (HHS), which is caused by a novel fowl adenovirus serotype 4 (FAdV-4), have been reported in chickens in parts of China, thereby causing huge economic losses to the poultry industry. Thus, an effective vaccine to control the further spread of infections with this hyper-virulent FAdV-4 is imperative. In this study, we isolated a novel FAdV-4 strain SDJN0105 from a broiler farm with HHS disease in Shandong Province. Pathogenicity was evaluated by the observation of clinical symptoms, necropsy changes, and pathological tissue sections after oral and intramuscular (IM) infection of Specific pathogen free (SPF) chickens. The chickens infected by IM injection all died within three days, and chickens infected via the oculonasal route died within five days post-infection (dpi). Histopathological examination revealed that the pathology was confined to heart, liver, spleen, lung, kidney, and particularly the liver. Irrespective of the inoculation route, the highest viral DNA copy numbers were detected in the livers of infected chickens. The mRNA expression levels of IL-1β, IL-6, IL-8, IFNs, TNF-α, Mx, and OASL were significantly upregulated during the viral infection. In addition, an inactivated oil-emulsion FAdV-4 vaccine was developed. The vaccine could provide full protection for SPF chickens against a lethal dose of the FAdV-4 strain SDJN0105 and a high level of antibodies. These results improve our understanding of the innate immune responses in chickens infected with FAdV-4 and the pathogenesis of FAdV-4 caused by host factors, and the developed FAdV-4 vaccine is promising as a drug candidate for the prevention and reduction of the spread of HHS in poultry in China.

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