Lateral Flow Assay for the Detection of African Swine Fever Virus Antibodies Using Gold Nanoparticle-Labeled Acid-Treated p72

Zhu, Wenzhuang; Meng, Kaiwen; Zhang, Yueping; Bu, Zhigao; Zhao, Dongming; Meng, Geng

doi:10.3389/fchem.2021.804981

Cited by 28 publications

(14 citation statements)

References 26 publications

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“…However, the p72 was shown to fail in forming trimers when expressed alone (Cobbold et al 2001 ). Interestingly, Zhu et al reported to express the 72 alone in yeast competent cells and obtain p72 trimers (Zhu et al 2022 ). In fact, the pB602L, an ASFV non-structural protein, has been described as a chaperone facilitating the correct folding of the p72 (Cobbold et al 2001 ; Epifano et al 2006 ; Meng et al 2020 ).…”

Section: Discussionmentioning

confidence: 99%

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Development of a p72 trimer–based colloidal gold strip for detection of antibodies against African swine fever virus

Geng

Sun

et al. 2022

Appl Microbiol Biotechnol

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African swine fever virus (ASFV) causes a highly contagious and often lethal swine viral disease, and leads to tremendous economic losses to the swine industry. Unfortunately, there are no vaccines and effective antiviral agents available to prevent and control ASFV outbreaks. Therefore, it is necessary to develop simple and rapid strategies to monitor ASFV-infected pigs to restrain its spread. In the current study, ASFV capsid protein p72 was expressed along with its chaperone pB602L to form trimers in human embryonic kidney 293 (HEK293) cells. The p72 trimers were subsequently labeled with colloidal gold to develop a immunochromatographic strip. The strip showed high specificity to ASFV-positive serum and no cross-reactivity to other swine virus positive sera. Importantly, the strip showed a higher sensitivity of detecting ASFV antibodies in both positive standard serum and clinical serum samples than a commercial enzyme-linked immunosorbent assay (ELISA) kit. Taken together, these results demonstrate the strip as a reliable diagnostic tool against ASFV infection, which will be appropriate for application in prevention and control of ASFV. Key points • ASFV p72 trimers were successfully generated . • A colloidal gold strip was developed based on ASFV p72 trimers . • The strip is appropriate for detecting ASFV antibodies in the field .

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Section: Discussionmentioning

confidence: 99%

“…Zhu et al have also utilized gold nanoparticle–labeled and acid-treated p72 to establish a lateral flow strip. The sensitivity of the strip is 1:10,000 for clinical positive sera (Zhu et al 2022 ). The sensitivity of our strip is 1:204,800, which is more sensitive than the commercial ELISA kit when testing positive standard serum.…”

Section: Discussionmentioning

confidence: 99%

Development of a p72 trimer–based colloidal gold strip for detection of antibodies against African swine fever virus

Geng

Sun

et al. 2022

Appl Microbiol Biotechnol

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“…Although ASF outbreaks had previously been limited in sub-Saharan Africa, an outbreak in Georgia in 2007 and its subsequent spread to neighboring countries in the south Caucasus, Europe, and the Russian Federation has highlighted the threat of ASF’s transboundary spread (Rowlands et al ., 2008; Sánchez-Vizcaíno et al ., 2015; Sauter-Louis et al ., 2021). Furthermore, ASF invaded China, the world’s largest pig producer in 2018, and then Southeast Asian countries, including Vietnam, continuing to have a devastating impact on the livestock industry and food security (Blome et al ., 2020; Le et al ., 2019; Wang et al ., 2020; WOAH, 2020; You et al ., 2021; Zhou et al ., 2018; Zhu et al ., 2022). As a result of the expansion of ASF, the pig industry has been suffering significant economic losses in ASF-endemic countries (Carriquiry et al ., 2020; Mason-D’Croz et al ., 2020; Nguyen-Thi et al ., 2021; WOAH, 2020; You et al ., 2021).…”

Section: Introductionmentioning

confidence: 99%

Development of a simple and highly sensitive method for the concentration and detection of African swine fever virus in oral fluids and its feasibility assessment on raised pigs in Northern Vietnam

Mai Thi,

Tran Thi Huong,

Dong Van

et al. 2024

Preprint

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Background: Early detection and early slaughter through quarantine are essential to prevent the spread of the African swine fever virus (ASFV). Highly accurate testing is effective for early detection, but it is still difficult to establish a system, especially in the Global South. Pooled oral fluids tests have been used for simple pathogen monitoring, but compared to blood tests, the virus concentration in oral fluids is low, resulting in false negative and missing true positive cases. We collected oral fluids from raised pigs in northern Vietnam and attempted a highly sensitive ASFV survey using a newly developed virus concentration and detection method. Results: In a spike test, the developed method showed up to 100 times greater sensitivity than a reference method. To compare and evaluate the performance of the developed method, a total of 68 pooled oral fluid samples were collected, 63 from northern Vietnam and 5 from southern Japan. Using real-time PCR, 9/68 (13.2%) were positive by the reference method, and 23/68 (33.8%) by the developed method. Using real-time LAMP, 1/68 (1.5%) were positive by the reference method and 6/68 (8.8%) by the developed method. Conclusions: The developed method improved the sensitivity of ASFV detection from oral fluids and enabled early diagnosis of pigs before the onset of the disease. The developed method has the potential to enable simple and highly sensitive diagnosis of ASF, which is essential for its early diagnosis and effective surveillance.

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“…Early pathogen detection is important when a disease lacks a vaccine as part of its prevention strategy. Thus, researchers have attempted to develop various serological diagnosis kits by using monoclonal antibodies (mAbs) against the ASFV immunogenic proteins such as [ 17 , 18 ] p. 72, [ 19 ] p. 62, [ 20 , 21 , 22 , 23 ] p. 54, [ 22 , 24 ] p. 30 , [ 25 ] p. 22 and others.…”

Section: Introductionmentioning

confidence: 99%

Novel Epitopes Mapping of African Swine Fever Virus CP312R Protein Using Monoclonal Antibodies

Hagoss

Shen

Zhang

et al. 2023

Viruses

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African Swine Fever (ASF) is a highly contagious and lethal pig disease and poses a huge threat to the pig industry worldwide. ASF virus (ASFV) encodes more than 150 different proteins, but the biological properties of most viral proteins are still unknown. ASFV CP312R protein has been proven to be one of the most immunogenic proteins during ASFV infection in pigs; however, its specific epitopes have yet to be identified. In this study, we verified the immunogenicity of CP312R protein in the sera from attenuated ASFV-inoculated pigs. We generated seven anti-ASFV CP312R mouse monoclonal antibodies (mAbs) from mice immunized with recombinant CP312R protein (rCP312R). All seven mAbs are the IgG2b-Kappa isotype and specifically interacted with the CP312R protein expressed in various cells that were infected by ASFVs or transfected with plasmid CP312R. The epitope mapping was performed by using these characterized mAbs and the peptide scanning (Pepscan) method followed by Western blot. As a result, two antigenic determinant regions were identified: two of the seven mAbs recognized the 122KNEQGEEIYP131 amino acids, and the remaining five mAbs recognized the 78DEEVIRMNAE87 amino acids of the CP312R protein. These antigenic determinants of CP312R are conserved in different ASFV strains of seven genotypes. By using the characterized mAb, confocal microscopy observation revealed that the CP312R was mainly localized in the cytoplasm and, to some extent, in nuclei and on the nuclear membrane of infected host cells. In summary, our results benefit our understanding on the antigenic regions of ASFV CP312R and help to develop better serological diagnosis of ASF and vaccine research.

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Lateral Flow Assay for the Detection of African Swine Fever Virus Antibodies Using Gold Nanoparticle-Labeled Acid-Treated p72

Cited by 28 publications

References 26 publications

Development of a p72 trimer–based colloidal gold strip for detection of antibodies against African swine fever virus

Development of a p72 trimer–based colloidal gold strip for detection of antibodies against African swine fever virus

Development of a simple and highly sensitive method for the concentration and detection of African swine fever virus in oral fluids and its feasibility assessment on raised pigs in Northern Vietnam

Novel Epitopes Mapping of African Swine Fever Virus CP312R Protein Using Monoclonal Antibodies

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