scholarly journals Prevalence and Genetic Analysis of Porcine Circovirus 3 in China From 2019 to 2020

2021 ◽  
Vol 8 ◽  
Author(s):  
Meng Ge ◽  
Jie Ren ◽  
Yi-Lin Xie ◽  
Dun Zhao ◽  
Fang-Cheng Fan ◽  
...  
Keyword(s):  
Potential Effect ◽  
Porcine Circovirus Type ◽  
Porcine Circovirus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Reading Frame ◽  
Humoral Immune ◽  
Antibody Positivity ◽  
Repeated Infections ◽  
Type 3

Porcine circovirus type 3 (PCV3), a virus belonging to the Circoviridae family, is considered to be associated with respiratory and neurological signs, cardiac and multisystemic inflammation, reproductive failure, and porcine dermatitis and nephropathy syndrome-like disease in pigs (Sus scrofa). In this study, epidemiological and serological investigations of PCV3 in clinically healthy pigs from different regions of China were performed. Overall, 42.87% (1,101/2,568) of pigs were positive for PCV3 Cap antibody via indirect enzyme-linked immunosorbent assay, with a higher prevalence of PCV3 in multiparous sows (62.22%, 881/1,416) and fattening pigs (28.96%, 159/549) than in suckling piglets (8.96%, 32/357) and nursery pigs (11.79%, 29/246). Of the 2,568 samples, 255 were further tested for PCV3 DNA using real-time polymerase chain reaction, and 63.14% of these were positive, with nearly half having <10 virus copies. The PCV3 DNA and antibody positivity rates were high in the pig serum samples; however, the virus titers and antibody levels were both low, indicating that the humoral immune response of PCV3-infected pigs was weak or lagging, and persistent or repeated infections could occur. Additionally, the complete genomes of 23 PCV3 strains were sequenced and analyzed, which showed nucleotide identities of 98.5~100.0%, 98.6~100.0%, and 99.2~100.0% in the complete genome, open reading frame (ORF)2, and ORF1 sequences, respectively, and amino acid identities of 96.7~100.0% and 99.3~100.0% in the capsid and replicase proteins, respectively. Phylogenetic analysis based on ORF2 nucleotide sequences indicated that the PCV3 strains obtained in the present study could be classified into three sub-clades, with most strains clustered into clade 3c, indicating that PCV3c is the dominant subtype in the regions of China investigated. In general, the present study revealed a high prevalence and high genetic divergence of PCV3 among Chinese pig herds, and indicated that the potential effect of PCV3 on the pig industry may be a concern.

scholarly journals Identification of porcine circovirus type 2 (PCV2), type 3 (PCV3) and porcine parvovirus (PPV) in swine by multiplex PCR test

2021 ◽  
Vol 20 (03) ◽  
pp. 11-17
Author(s):  
Phat X. Dinh
Keyword(s):  
Multiplex Pcr ◽  
Real Time ◽  
Porcine Circovirus Type ◽  
Porcine Circovirus Type 2 ◽  
Porcine Circovirus ◽  
Serum Samples ◽  
Single Target ◽  
Ns1 Gene ◽  
Type 3

This study aimed to simultaneously detect three important viruses reported to be involved in the reproductive problems of sows. A multiplex PCR (mPCR) test was developed to provide rapid diagnosis of porcine circovirus type 2 and 3 (PCV2, PCV3) and to illustrate parvovirus (PPV) prevalence in sow herds. Three pairs of specific primers were designed to target PCV2 Cap gene, PCV3 Cap gene and PPV NS1 gene, with predicted mPCR products of 702 bp, 267 bp and 380 bp, respectively. The detection limit of mPCR was 100 copies/reaction per target gene. The mPCR was run against a panel of 94 swine serum samples whose infection status had been pre-determined by commercial real-time PCR kits. Sequencing of mPCR products performed with clinical serum samples accurately confirmed the results. Overall, the results indicated that the mPCR functioned accurately and specifically and matched 100% with the single-target real-time PCRs. The mPCR was developed successfully and can be used in routine diagnosis of PCV2, PCV3 and PPV.

scholarly journals Identification of Porcine circovirus type (PCV2) and type 3 (PCV3), Porcine 2 parvovirus (PPV) in swine by multiplex PCR test

2020 ◽  
Author(s):  
Thoai Kim Tran ◽  
Trang Thi Thanh Nguyen ◽  
Hiep Lai Xuan Vu ◽  
Phat Xuan Dinh
Keyword(s):  
Multiplex Pcr ◽  
Real Time ◽  
Porcine Circovirus Type ◽  
Rapid Diagnosis ◽  
Porcine Circovirus ◽  
Serum Samples ◽  
Ns1 Gene ◽  
Type 3 ◽  
Swine Serum

Abstract Background : Aiming to simultaneously detect three important viruses known to be involved in reproductive problems of sows, a multiplex PCR (mPCR) test was developed to provide rapid diagnosis of porcine circovirus type 2 and 3 (PCV2, PCV3) and to illustrate parvovirus (PPV) prevalence in sow herds. Methods : Three pairs of specific primers were designed to target PCV2 Cap gene, PCV3 Cap gene and PPV NS1 gene, with predicted mPCR products of 702 bp, 267 bp and 380 bp, respectively. Results : The detection limit of mPCR was 100 copies/ reaction per target gene. Sequencing of mPCR products performed with clinical serum samples accurately confirmed results. The mPCR was run against a panel of 94 swine serum samples whose infection status had been pre-determined by commercial real-time PCR kits. Overall, the mPCR results matched 100% with the real-time PCRs. Conclusions : The developed mPCR test functions successfully and can be used in routine rapid diagnosis of PCV2, PCV3 and PPV.

The presence and genetic characteristics of porcine circovirus 3 from pigs in Southern and Central provinces of Vietnam

2020 ◽  
pp. 26-31
Author(s):  
Giao N. P. Trinh
Keyword(s):  
Gene Segment ◽  
Porcine Circovirus Type ◽  
Porcine Circovirus ◽  
Nucleotide Sequencing ◽  
Duplex Pcr ◽  
Serum Samples ◽  
Swine Industry ◽  
Genetic Characteristics ◽  
Swine Herds ◽  
Type 3

Porcine circovirus type 3 (PCV3) is an emerging circovirus species that has recently been reported in different countries around the world, suggesting a widespread circulation. This study was carried out in order to investigate the presence and further genetic characteristics of PCV3 from swine herds in Southern and Central provinces of Vietnam. A duplex PCR assay for rapid detection of PCV3 in pigs was established with a pair of specific primers designed between rep and cap gene segment to amplify full-length ORF2 and another set of primers binding to COX1gene serving as an internal amplification control (IAC). The resulting duplex PCR was used to examine PCV3 presence in 94tissue and serum samples. Subsequently, PCV3 was detected in 10 out of 94 cases (10.6%). The infection rate in sows (14.3%) was higher than that in grower pigs (7.7%). Regarding nucleotide sequence comparison, 10 ORF2 genes were selected for nucleotide sequencing and their alignment showed 97.2% - 99.5% homology. According to the phylogenetic analysis and sequence alignment of cap gene, all the sequences were clustered into group PCV3a,including 9 strains of sub-group PCV3a1 and only one strain of subgroup PCV3a2. These findings indicated that the PCV3a group is circulating in swine farms in Vietnam. This study provides better insights into epidemiology of this pathogen in the national swine industry.

scholarly journals A Molecular Survey and Phylogenetic Analysis of Porcine Circovirus Type 3 Using Oral Fluid, Faeces and Serum

2020 ◽  
Author(s):  
Jan Plut ◽  
Urska Jamnikar-Ciglenecki ◽  
Irena Golinar-Oven ◽  
Tanja Knific ◽  
Marina Stukelj
Keyword(s):  
Genome Sequencing ◽  
Oral Fluid ◽  
Statistical Significance ◽  
Age Groups ◽  
Porcine Circovirus Type ◽  
Porcine Circovirus Type 2 ◽  
Porcine Circovirus ◽  
Serum Samples ◽  
Type 3

Abstract Background: Porcine circovirus type 3 is the most recently discovered porcine circovirus, and an emerging pathogen. In this study the status of its presence on some Slovenian farms is reported. The effectiveness of the vaccine against porcine circovirus type 2 was assessed against porcine circovirus type 3.Group samples of oral fluid, faeces and individual serum samples were taken from six different pig categories and tested for presence of viral DNA, using both real time and conventional PCR. Positive samples were subjected to direct Sanger sequencing. Nucleotide sequences were analyzed and compared to GenBank PCV3 sequences.Results: Positive samples were sent for genome sequencing, which confirmed the presence of virus in all different pig categories on five farms. A high to moderate correlation of strong statistical significance was found between individual serum samples, oral fluid and faeces. Slovenian PCV3 was found to be distributed in a way similar to that of other countries. Slovenian PCV3 nt sequences are highly related, sharing more than 99.5 % nt identity. On one farm a commercially available vaccine against porcine circovirus type 2 was used on 3-week-old pigs. It did not affect the presence of porcine circovirus type 3 in oral fluid or sera of any of the seven age groups of pigs, each with two control groups.Conclusions: The results constitute the first discovery of the virus in Slovenia. Genome sequencing has revealed a high degree of similarity between Slovenian and GenBank isolates.

scholarly journals Maternally Derived Antibody Levels Influence on Vaccine Protection against PCV2d Challenge

Animals ◽  
2021 ◽  
Vol 11 (8) ◽  
pp. 2231
Author(s):  
István Kiss ◽  
Krisztina Szigeti ◽  
Zalán G. Homonnay ◽  
Vivien Tamás ◽  
Han Smits ◽  
...  
Keyword(s):  
Immune Response ◽  
Humoral Immune Response ◽  
Subunit Vaccine ◽  
Porcine Circovirus Type ◽  
Porcine Circovirus ◽  
Vaccine Protection ◽  
Humoral Immune ◽  
Negative Effect ◽  
Antibody Levels

Piglets from a porcine circovirus type 2 (PCV2) stable farm of low and high levels of maternally derived antibodies (MDA) against PCV2 were vaccinated either with a whole virus type or a PCV2 ORF2 antigen-based commercial subunit vaccine at three weeks of age. Two non-vaccinated groups served as low and high MDA positive controls. At four weeks post vaccination, all piglets were challenged with a PCV2d-2 type virus strain and were checked for parameters related to vaccine protection over a four-week observation period. MDA levels evidently impacted the outcome of the PCV2d-2 challenge in non-vaccinated animals, while it did not have a significant effect on vaccine-induced protection levels. The humoral immune response developed faster in the whole virus vaccinates than in the subunit vaccinated pigs in the low MDA groups. Further, high MDA levels elicited a stronger negative effect on the vaccine-induced humoral immune response for the subunit vaccine than for the whole virus vaccine. The group-based oral fluid samples and the group mean viraemia and faecal shedding data correlated well, enabling this simple, and animal welfare-friendly sampling method for the evaluation of the PCV2 viral load status of these nursery piglets.

scholarly journals Epidemiological investigation of porcine circovirus type 2 and its coinfection rate in Shandong province in China from 2015 to 2018

2021 ◽  
Vol 17 (1) ◽  
Author(s):  
Zicheng Ma ◽  
Mengda Liu ◽  
Zhaohu Liu ◽  
Fanliang Meng ◽  
Hongyu Wang ◽  
...  
Keyword(s):  
Pseudorabies Virus ◽  
Porcine Circovirus Type ◽  
Porcine Circovirus Type 2 ◽  
Shandong Province ◽  
Porcine Circovirus ◽  
Respiratory Syndrome Virus ◽  
Limited Information ◽  
Serum Samples ◽  
Diarrhea Virus

Abstract Background Porcine circovirus type 2 (PCV2) is one of the crucial swine viral pathogens, caused porcine circovirus associated diseases (PCVAD). Shandong province is one of the most important pork producing areas and bears a considerable economic loss due to PCVAD. However, there is limited information on epidemiology and coinfection rate of PCV2 with other critical swine diseases in this area, such as porcine reproductive and respiratory syndrome virus (PRRSV), classical swine fever virus (CSFV), Pseudorabies virus (PRV), and porcine epidemic diarrhea virus (PEDV). Results Overall, 89.59% serum samples and 36.98% tissue samples were positive for PCV2 specified ELISA and PCR positive for PCV2, respectively. The coinfection rates of PCV2 with PRRSV, PRV, CSFV, and PEDV were 26.73%, 18.37%, 13.06%, and 3.47%, respectively. Moreover, genetic characteristic of PCV2 were analyzed based on the cap genes showing that PCV2d is the dominant sub-genotype circulating in the province. Conclusions Our findings reveal that PCV2d, as the dominant strain, is prevailing in pig farms in Shandong province at high levels. There was a high frequency of coinfection of PCV2 and PRRSV.

scholarly journals Purification of Porcine Circovirus Type 2 Using an Affinity Chromatography Based on a Neutralizing Monoclonal Antibody against Viral Capsid Protein

Pathogens ◽  
2021 ◽  
Vol 10 (12) ◽  
pp. 1564
Author(s):  
Haiqiao Bian ◽  
Chong Yu ◽  
Yanwu Wei ◽  
Li Feng ◽  
Changming Liu ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Monoclonal Antibody ◽  
Affinity Chromatography ◽  
Porcine Circovirus Type ◽  
Porcine Circovirus Type 2 ◽  
Porcine Circovirus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Chromatography Method ◽  
Viral Adsorption

Porcine circovirus type 2 (PCV2) is a DNA virus without an envelope. The viral particle is icosahedral and has a diameter of approximately 17 nm. In order to obtain the purified virus, a broad-spectrum monoclonal antibody 3A5 against PCV2 was coupled to CNBr-activated SepharoseTM 4B, and an affinity chromatography was established for PCV2 purification. A total of 6.5 mg of purified PCV2a/LG with 97% purity was obtained from 120 mL of the viral culture medium, and only PCV2 was detected by electron microscopy. No significant changes in the antigenic characteristics of the purified virus were detected by a capture enzyme-linked immunosorbent assay (ELISA). Furthermore, the titer of the purified PCV2 was 100 times higher than that of the unpurified virus. This affinity chromatography method was also used to purify PCV2b/LN590516 and PCV2d/SD446F16, and the purified viruses were detected by electron microscopy, capture ELISA, and virus titration, respectively. The results showed that these two strains can be successfully purified, but the yield is lower than that of the PCV2a strain. In addition, the purified virus could be used to study the viral adsorption and invasion of PK15 cells using indirect immunofluorescence assays. A large number of PCV2 signals were detected to transfer from the cellular surface to the periphery of the nucleus of the PK15 cells after 30 min of adsorption of the PCV2 to the PK15 cells. The affinity chromatography is a simple and convenient tool to obtain PCV2 with high purity. It could be applied for virus structure analysis, antibody preparation, and viral adsorption and invasion research.

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