Development of a real‐time PCR assay for detection of African swine fever virus with an endogenous internal control

Wang, Yin; Xu, Lizhe; Noll, Lance; Stoy, Colin; Porter, Elizabeth; Fu, Jinping; Yan, Feng; Peddireddi, Lalitha; Liu, Xuming; Dodd, Kimberly A.; Jia, Wei Hua; Bai, Jianfa

doi:10.1111/tbed.13582

Cited by 60 publications

(38 citation statements)

References 23 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Polymerase chain reaction protocols represent the first line of ASFV detection, and an increasing number of published protocols and fully validated test kits are available [ 196 , 197 , 198 , 199 , 200 , 201 , 202 , 203 , 204 , 205 , 206 , 207 , 208 ]. Regarding the later, commercial kits with reasonable pricing, the integration of internal control systems and the lack of need for extra consumables are key.…”

Section: Diagnosis Of Asf In Wild Boarmentioning

confidence: 99%

African Swine Fever in Wild Boar in Europe—A Review

Sauter‐Louis

Conraths

Probst

et al. 2021

Viruses

110

114

View full text Add to dashboard Cite

The introduction of genotype II African swine fever (ASF) virus, presumably from Africa into Georgia in 2007, and its continuous spread through Europe and Asia as a panzootic disease of suids, continues to have a huge socio-economic impact. ASF is characterized by hemorrhagic fever leading to a high case/fatality ratio in pigs. In Europe, wild boar are especially affected. This review summarizes the currently available knowledge on ASF in wild boar in Europe. The current ASF panzootic is characterized by self-sustaining cycles of infection in the wild boar population. Spill-over and spill-back events occur from wild boar to domestic pigs and vice versa. The social structure of wild boar populations and the spatial behavior of the animals, a variety of ASF virus (ASFV) transmission mechanisms and persistence in the environment complicate the modeling of the disease. Control measures focus on the detection and removal of wild boar carcasses, in which ASFV can remain infectious for months. Further measures include the reduction in wild boar density and the limitation of wild boar movements through fences. Using these measures, the Czech Republic and Belgium succeeded in eliminating ASF in their territories, while the disease spread in others. So far, no vaccine is available to protect wild boar or domestic pigs reliably against ASF.

show abstract

Section: Diagnosis Of Asf In Wild Boarmentioning

confidence: 99%

African Swine Fever in Wild Boar in Europe—A Review

Sauter‐Louis

Conraths

Probst

et al. 2021

Viruses

110

114

View full text Add to dashboard Cite

show abstract

“…In silico analysis showed the PCR set can perfectly match 75.3 % of PCV2a strains, 86.3 % of PCV2b strains, and 78.1 % of PCV2d strains. Assuming a single nucleotide mismatch to a primer (excluding the more critical 5 bp in the 3′ end) or a probe (excluding the more critical 5 bp in the middle) in the assay can still amplify and generate signal ( Wang et al, 2020b ), the strain coverages would be increased to 93.4 %, 95.1 % and 93.6 % for the three genotypes, respectively. Information of all primers and probes are shown in Table 1 .…”

Section: Resultsmentioning

confidence: 99%

Development of a differential multiplex real-time PCR assay for porcine circovirus type 2 (PCV2) genotypes PCV2a, PCV2b and PCV2d

Wang

Noll

Porter

et al. 2020

Journal of Virological Methods

Self Cite

View full text Add to dashboard Cite

Highlights A multiplex real-time PCR (mqPCR) is developed for PCV2a, 2b, and 2d differentiations. Sequencing of 74 strains confirmed genotyping results generated by the mqPCR. The mqPCR did not detect any non-target swine pathogens included in this study. This genotyping assay is recommended following a general PCV2 diagnostic testing.

show abstract

“…Thus, it is essential to differentiate the wide-type ASFV from vaccines with gene deleted for preventing and controlling ASF. However, most of the existing detection methods, including PCR, real-time PCR, LAMP assay and RPA assay, were developed and improved based on the conserved region of B646L gene, which can not apply to the detection of gene deletions strains [2,16,17]. In this study, we developed and evaluated a duplex real-time PCR targeting the B646L gene and MGF505-2R gene.…”

Section: Discussionmentioning

confidence: 99%

Development and Evaluation of Duplex TaqMan Real-Time PCR Assay for Detection and Differentiation of Wide-Type and MGF505 Gene-Deleted African Swine Fever Viruses

Guo

Li²,

Qiao

et al. 2020

Preprint

View full text Add to dashboard Cite

Background: African swine fever (ASF) is the most important disease to the pigs and cause serious economic losses to the countries with large-scale swine production. Vaccines are recognized as the most useful tool to prevent and control ASF virus (ASFV) infection. Currently, the MGF505 and MGF360 gene-deleted ASFVs or combined with CD2v deletion were confirmed to be the most promising vaccine candidates. Thus, it is essential to develop a diagnosis method to discriminate wide-type strain from the vaccines used. Results: In this study, we established a duplex TaqMan real-time PCR based on the B646L gene and MGF505-2R gene. The sequence alignment showed that the targeted regions of primers and probes are highly conserved in the genotype II ASFVs. The duplex real-time assay can specifically detect B646L and MGF505-2R gene single or simultaneously without cross-reaction with other porcine viruses tested. The limit of detection was 5.8 copies and 3.0 copies for the standard plasmids containing B646L and MGF505-2R genes, respectively. Clinical samples were tested in parallel by duplex real-time PCR and a commercial ASFV detection kit. The detection results of these two assays against B646L gene were well consistent. Conclusion: We successfully developed and evaluated a duplex TaqMan real-time PCR method which can effectively distinguish the wide type and MGF505 gene-deleted ASFVs. It would be a useful tool for the clinical diagnosis and control of ASF.

show abstract

Development of a real‐time PCR assay for detection of African swine fever virus with an endogenous internal control

Cited by 60 publications

References 23 publications

African Swine Fever in Wild Boar in Europe—A Review

African Swine Fever in Wild Boar in Europe—A Review

Development of a differential multiplex real-time PCR assay for porcine circovirus type 2 (PCV2) genotypes PCV2a, PCV2b and PCV2d

Development and Evaluation of Duplex TaqMan Real-Time PCR Assay for Detection and Differentiation of Wide-Type and MGF505 Gene-Deleted African Swine Fever Viruses

Contact Info

Product

Resources

About