Antigenic Regions of African Swine Fever Virus Phosphoprotein P30

Wu, Ping; Lowe, Anthony D.; Rodriguez, Yelitza Y.; Murgia, Maria V.; Dodd, Kimberly A.; Rowland, Raymond R.R.; Jia, Wei Hua

doi:10.1111/tbed.13533

Cited by 18 publications

(17 citation statements)

References 64 publications

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“…The expressed CD2v protein can be used to study the interaction between receptors and ligand, the process of which requires further investigation. ASFV p30 is a phosphorylated protein expressed at the early stage in infected cells [18]. ELISA kits based on p30 are widely used to detect ASFV infection [18][19][20].…”

Section: Discussionmentioning

confidence: 99%

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Development of a Dual ELISA for the Detection of CD2v-Unexpressed Lower-Virulence Mutational ASFV

Zhao

Jiang

et al. 2021

Life

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African swine fever virus (ASFV) is an important viral pathogen infecting pigs worldwide throughout the pig industry. CD2v (an outer-membrane glycosylated protein of ASFV)-unexpressed lower-virulence mutants have appeared in China and other countries in recent years. Using OIE-recommended quantitative PCR and ELISA methods, people can accurately judge whether pigs are infected with wild-type ASFV. However, the strategy has failed to distinguish ΔCD2v lower-virulence mutants and wild-type ASFV infection. Here, we expressed and purified the CD2v and p30 proteins via CHO cells and successfully established a dual enzyme-linked immunosorbent assay (ELISA), which can be used to differentiate pigs infected with wild-type ASFV or with CD2v-unexpressed lower-virulence mutants. The dual ELISA showed excellent specificity without cross-reactions with antibodies of PRRSV, CSFV, JEV, PRV, or PPV. The dual ELISA could detect ASFV-infected positive serum samples up to dilutions of 5120 times, possessing high sensitivity. Therefore, the application of this dual ELISA approach can play an important role in ASFV epidemiology study and fill the gaps in differential diagnosis.

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Section: Discussionmentioning

confidence: 99%

“…ASFV p30 is a phosphorylated protein expressed at the early stage in infected cells [18]. ELISA kits based on p30 are widely used to detect ASFV infection [18][19][20]. In this study, the ELISA kit based…”

Section: Discussionmentioning

confidence: 99%

Development of a Dual ELISA for the Detection of CD2v-Unexpressed Lower-Virulence Mutational ASFV

Zhao

Jiang

et al. 2021

Life

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“…There are significant differences in the amino acid composition and identity of the new field strain compared to some other members of genotype I. The dissimilarity in the amino acid composition could be due to amino acid substitution [ 29 ] or mutation. Changes may result from vaccination of pig herds with classical swine fever vaccines by some medium- and large-scale pig farmers in Southwest Nigeria, as observed in earlier epizootic [ 1 ].…”

Section: Discussionmentioning

confidence: 99%

Detection of African swine fever virus in pigs in Southwest Nigeria

et al. 2021

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Background and Aim: Nigeria experienced repeated outbreaks of African swine fever (ASF) in pig herds between 1997 and 2005 in the southwest region of the country. ASF is believed to currently be enzootic in this region. The status of enzootic transmission of ASF virus strain to pigs is; however, unknown. Twenty-three genotypes of the ASF virus based on the p72 gene are found across Africa. This study aimed to identify the current circulating field strain(s) of the ASF virus in Southwest Nigeria and characterized evolutionary trends. Materials and Methods: DNA samples were extracted from 144 pooled blood samples obtained from 2012 to 2013 following the manufacturer's instructions. DNA was used for conventional polymerase chain reaction using primers targeting the p72 gene and amplified products sequenced with Sanger's sequencing. Sequences were analyzed for homology and phylogenetic relationships. Results: Eleven of 144 samples (7.6%) showed bands at 950 bp. A new field strain of ASF virus of genotype I that shared ancestry with ASF virus strains or isolates from Spain and Brazil was identified among pig herds. The new strain differs phylogenetically in amino acid composition compared with previously identified ASF virus field strains. Conclusion: The currently circulating field strain of ASF virus suggests a mutation responsible for decreased morbidity and mortality recorded in sporadic cases.

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“…Bootstrap values were calculated based on 1000 replicates by the neighbor-joining method. The immunogenic proteins p30 and p54 were also further analyzed to determine whether there were any changes in the antigenic regions of ASFV among different culture passages and references [17,18].…”

Section: Sequencing and Genetic Analysis In Serial Passagesmentioning

confidence: 99%

Development of Optimized Protocol for Culturing African Swine Fever Virus (ASFV) Field Isolates in MA104 Cells

Kwon

Tien²,

Vo³

et al. 2021

Preprint

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I. Background: ASFV causes a highly contagious hemorrhagic disease with a high mortality rates in domestic pigs. The virus has been isolated across various cell lines, but identifying a cell line to develop an effective commercial vaccine has been challenging which a major obstacle to effective vaccine development is identifying a commercial cell line that is suitable for high-yield viral replication.II. Methods and Results: The goal of this study was to identify a candidate commercial cell line for the replication of African swine fever virus (ASFV) by comparing several available cell lines with various medium factors. In the sensitivity test of cells, MA104 and MARC-145 had strong potential for ASFV replication. Next, MA104 cells were used to compare the adaptation of ASFV obtained from tissue homogenates and blood samples in various infectious media. At the 10th passage, the ASFV obtained from the blood sample had a significantly higher viral load than that obtained from the tissue sample (P = 0.000), exhibiting a mean Ct value = 20.39 ± 1.99 compared with 25.36 ± 2.11. For blood samples, ASFV grew on infectious medium B more robustly than on infectious medium A (P = 0.006), corresponding to a Ct value = 19.58 ± 2.10 versus 21.20 ± 1.47. ASFV originating from blood specimens continued to multiply gradually and peaked in the 15th passage, exhibiting a Ct value = 14.36 ± 0.22 in infectious medium B and a Ct value = 15.42 ± 0.14 in infectious medium A. However, there was no difference (P = 0.062) in ASFV growth between infectious media A and B when ASFV was cultured from tissue homogenates. III. Conclusions: A model was developed to enhance ASFV replication through adaptation to MA104 cells and the lack of mutation in serial culture passages may serve to maintain the immunogenicity of ASFV isolates when they are developed as vaccine candidates.

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Antigenic Regions of African Swine Fever Virus Phosphoprotein P30

Cited by 18 publications

References 64 publications

Development of a Dual ELISA for the Detection of CD2v-Unexpressed Lower-Virulence Mutational ASFV

Development of a Dual ELISA for the Detection of CD2v-Unexpressed Lower-Virulence Mutational ASFV

Detection of African swine fever virus in pigs in Southwest Nigeria

Development of Optimized Protocol for Culturing African Swine Fever Virus (ASFV) Field Isolates in MA104 Cells

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