Development of Diagnostic Tests Provides Technical Support for the Control of African Swine Fever

Qiu, Zilong; Li, Zhaoyao; Yan, Quanhui; Li, Yuwan; WenJie, Xiong; Wu, Keke; Li, Xiaowen; Fan, Songqing; Zhao, Mingqiu; Ding, Hongxing; Chen, Jinding

doi:10.3390/vaccines9040343

Cited by 34 publications

(26 citation statements)

References 137 publications

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“…At present, based on CRISPR/Cas12a-based nucleic acid detection technology, recombinase polymerase amplification and gold nanoparticles technology have been reported for the rapid detection of ASFV. However, its commercial use still needs a particular time [ 33 – 36 ]. Currently widely used detection technology is the real-time PCR targeting the p72 gene, which shows high sensitivity and specificity.…”

Section: Strategies For Generating Asf Lav Candidatesmentioning

confidence: 99%

Current efforts towards safe and effective live attenuated vaccines against African swine fever: challenges and prospects

et al. 2021

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Background African swine fever (ASF) is a fatal hemorrhagic disease in domestic pigs and wild boar caused by African swine fever virus (ASFV). Since ASF has been introduced into Europe and Asia, the major pig-raising areas, posing a huge threat to the pork industry worldwide. Currently, prevention and control of ASF are basically dependent on strict biosecurity measures and stamping-out policy once ASF occurs. Main text The major risks of ASF spread are insufficient biosecurity measures and human behaviors. Therefore, a safe and effective vaccine seems to be a reasonable demand for the prevention and control of ASF. Due to the efficacy advantage over other types of vaccines, live attenuated vaccines (LAVs), especially virulence-associated genes deleted vaccines, are likely to be put into emergency and conditional use in restricted areas if ASF is out of control in a country with a huge pig population and pork consumption, like China. However, the safety, efficacy, and genetic stability of current candidate ASF LAVs require comprehensive clinical evaluations prior to country-wide field application. Several critical issues need to be addressed to commercialize an ideal ASF LAV, including a stable cell line for manufacturing vaccines, differentiation of infected from vaccinated animals (DIVA), and cross-protection from different genotypes. Conclusion A safe and effective DIVA vaccine and an accompanying diagnostic assay will facilitate the prevention, control, and eradication of ASF, which is quite challenging in the near future. Graphical Abstract

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Section: Strategies For Generating Asf Lav Candidatesmentioning

confidence: 99%

Current efforts towards safe and effective live attenuated vaccines against African swine fever: challenges and prospects

et al. 2021

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“…Moreover, it requires a large and well-established database to support the results, which have limited development in the field of rapid detection of viruses (Pelletier et al 2011). In addition, enzyme-linked immunosorbent assay (ELISA) is the most typical immunological method, which has been approved for a lot of diseases detection by FDA and OIE (Kazakova et al 2017;Qiu et al 2021;Zhang et al 2020). But many researchers had found that ELISA showed low sensitivity of 77.2% in the commercial kits, poor thermal stability, poor repeatability, and false positive due to the interference of autoantibodies (Gallardo et al 2015).…”

Section: Introductionmentioning

confidence: 99%

One-pot platform for rapid detecting virus utilizing recombinase polymerase amplification and CRISPR/Cas12a

Xiong

Cao

Chen

et al. 2022

Appl Microbiol Biotechnol

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The livestock industry has been deeply affected by African swine fever virus (ASFV) and Capripoxvirus (CaPV), which caused an enormous economic damage. It is emergent to develop a reliable detection method. Here, we developed a rapid, ultra-sensitive, and one-pot DNA detection method combining recombinase polymerase amplification (RPA) and CRISPR/Cas12a for ASFV and CaPV, named one-pot-RPA-Cas12a (OpRCas) platform. It had the virtue of both RPA and CRISPR/Cas12a, such as high amplification efficiency, constant temperature reaction, and strict target selectivity, which made diagnosis simplified, accurate and easy to be operated without expensive equipment. Meanwhile, the reagents of RPA and CRISPR/Cas12a were added to the lid and bottom of tube in one go, which overcame the incompatibility of two reactions and aerosol contamination. To save cost, we only need a quarter of the amount of regular RPA per reaction which is enough to achieve clinical diagnosis. The OpRCas platform was 10 to 100 times more sensitive than qPCR; the limit of detection (LOD) was as low as 1.2 × 10 −6 ng/µL (3.07 copies/µL by ddPCR) of ASFV and 7.7 × 10 −5 ng/ µL (1.02 copies/µL by ddPCR) of CaPV with the portable fluorometer in 40 min. In addition, the OpRCas platform combined with the lateral flow assay (LFA) strip to suit for point-of-care (POC) testing. It showed 93.3% consistency with qPCR for clinical sample analysis. Results prove that OpRCas platform is an easy-handling, ultra-sensitive, and rapid to achieve ASFV and CaPV POC testing. Key points• The platform realizes one-pot reaction of RPA and Cas12a.• Sensitivity is 100 times more than qPCR.• Three output modes are suitable to be used to quantitative test or POC testing.

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“…Strict biosecurity protocols and classic sanitary methods both at the farm and at national levels have been adopted to fight the spread of this virus; however, these strategies are remain insufficient, and in some countries they are difficult or impossible to follow owing to the lack of available resources [ 9 , 10 , 11 , 12 ]. As there are no licensed vaccines or treatments available, the most important control strategies for ASFV are early virus detection, followed by laboratory diagnosis [ 13 ], and the subsequent culling of ASFV-infected animals from the herds to prevent the spread of the virus [ 14 ].…”

Section: Introductionmentioning

confidence: 99%

Cell Lines for the Development of African Swine Fever Virus Vaccine Candidates: An Update

2022

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African swine fever virus (ASFV) is the etiological agent of a highly lethal disease in both domestic and wild pigs. The virus has rapidly spread worldwide and has no available licensed vaccine. An obstacle to the construction of a safe and efficient vaccine is the lack of a suitable cell line for ASFV isolation and propagation. Macrophages are the main targets for ASFV, and they have been widely used to study virus–host interactions; nevertheless, obtaining these cells is time-consuming and expensive, and they are not ethically suitable for the production of large-scale vaccines. To overcome these issues, different virulent field isolates have been adapted on monkey or human continuous cells lines; however, several culture passages often lead to significant genetic modifications and the loss of immunogenicity of the adapted strain. Thus, several groups have attempted to establish a porcine cell line able to sustain ASFV growth. Preliminary data suggested that some porcine continuous cell lines might be an alternative to primary macrophages for ASFV research and for large-scale vaccine production, although further studies are still needed. In this review, we summarize the research to investigate the most suitable cell line for ASFV isolation and propagation.

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Development of Diagnostic Tests Provides Technical Support for the Control of African Swine Fever

Cited by 34 publications

References 137 publications

Current efforts towards safe and effective live attenuated vaccines against African swine fever: challenges and prospects

Current efforts towards safe and effective live attenuated vaccines against African swine fever: challenges and prospects

One-pot platform for rapid detecting virus utilizing recombinase polymerase amplification and CRISPR/Cas12a

Cell Lines for the Development of African Swine Fever Virus Vaccine Candidates: An Update

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