Development of a Cleaved Probe-Based Loop-Mediated Isothermal Amplification Assay for Rapid Detection of African Swine Fever Virus

Wang, Songqi; Shen, Huiyong; Lin, Quankui; Huang, Jun; Zhang, Chunhong; Liu, Zhicheng; Sun, Mei; Zhang, Jianfeng; Liao, Ming; Li, Yugu; Zhang, Jianmin

doi:10.3389/fcimb.2022.884430

Cited by 4 publications

(3 citation statements)

References 23 publications

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“…This technique entails the design of four primers that specifically target six regions on the desired gene, and employs DNA polymerase for amplification under constant temperature conditions (118). As presented in Table 2, various LAMP detection methods have been developed to target specific viral genes such as K78R, B646L, 9GL, and TPII (61)(62)(63)(64), with LODs of 30 copies/μL, 10 copies/reaction, 13 copies/μL, and 400 copies/reaction, respectively. Wang et al has devised a ladder-shaped melting temperature isothermal amplification (LMTIA) technique that specifically targets the ASFV-B646L gene.…”

Section: Loop-mediated Isothermal Amplificationmentioning

confidence: 99%

Current detection methods of African swine fever virus

Hu,

Tian,

Lai

et al. 2023

Front. Vet. Sci.

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African swine fever (ASF), caused by the African swine fever virus (ASFV), is a highly contagious and notifiable animal disease in domestic pigs and wild boars, as designated by the World Organization for Animal Health (WOAH). The effective diagnosis of ASF holds great importance in promptly controlling its spread due to its increasing prevalence and the continuous emergence of variant strains. This paper offers a comprehensive review of the most common and up-to-date methods established for various genes/proteins associated with ASFV. The discussed methods primarily focus on the detection of viral genomes or particles, as well as the detection of ASFV associated antibodies. It is anticipated that this paper will serve as a reference for choosing appropriate diagnostic methods in diverse application scenarios, while also provide direction for the development of innovative technologies in the future.

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Section: Loop-mediated Isothermal Amplificationmentioning

confidence: 99%

Current detection methods of African swine fever virus

Hu,

Tian,

Lai

et al. 2023

Front. Vet. Sci.

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“…Non-specific fluorescent methods use molecules that are fluorescent only when double-stranded DNA is present [53]. Specific methods use nucleic acids tagged with fluorophores and quenchers that are specific for the nucleic acid target and fluoresce only after the successful amplification of that target [25,41,54,55]. Methods such as bioluminescent assay in real time (BART) require a luminometer to measure luminescence produced by luciferase [56].…”

Section: Poc Nucleic Acid Testsmentioning

confidence: 99%

Point-of-Care Testing for Hepatitis Viruses: A Growing Need

Pauly,

Ganova-Raeva

2023

Life

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Viral hepatitis, caused by hepatitis A virus (HAV), hepatitis B virus (HBV), hepatitis C virus (HCV), hepatitis D virus (HDV), or hepatitis E virus (HEV), is a major global public health problem. These viruses cause millions of infections each year, and chronic infections with HBV, HCV, or HDV can lead to severe liver complications; however, they are underdiagnosed. Achieving the World Health Organization’s viral hepatitis elimination goals by 2030 will require access to simpler, faster, and less expensive diagnostics. The development and implementation of point-of-care (POC) testing methods that can be performed outside of a laboratory for the diagnosis of viral hepatitis infections is a promising approach to facilitate and expedite WHO’s elimination targets. While a few markers of viral hepatitis are already available in POC formats, tests for additional markers or using novel technologies need to be developed and validated for clinical use. Potential methods and uses for the POC testing of antibodies, antigens, and nucleic acids that relate to the diagnosis, monitoring, or surveillance of viral hepatitis infections are discussed here. Unmet needs and areas where additional research is needed are also described.

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“…Therefore, after hybridization of the probe with the target sequence, ribonuclease H2 cleaves the probe, releasing the quencher and leading to the emission of a fluorescence signal. This assay has a detection limit of 13 copies/µL and provides results in 40 min [49].…”

Section: Loop-mediated Isothermal Amplification (Lamp)mentioning

confidence: 99%

Emergent Molecular Techniques Applied to the Detection of Porcine Viruses

Flores-Contreras,

Carrasco-González,

Linhares

et al. 2023

Veterinary Sciences

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Molecular diagnostic tests have evolved very rapidly in the field of human health, especially with the arrival of the recent pandemic caused by the SARS-CoV-2 virus. However, the animal sector is constantly neglected, even though accurate detection by molecular tools could represent economic advantages by preventing the spread of viruses. In this regard, the swine industry is of great interest. The main viruses that affect the swine industry are described in this review, including African swine fever virus (ASFV), porcine reproductive and respiratory syndrome virus (PRRSV), porcine epidemic diarrhea virus (PEDV), and porcine circovirus (PCV), which have been effectively detected by different molecular tools in recent times. Here, we describe the rationale of molecular techniques such as multiplex PCR, isothermal methods (LAMP, NASBA, RPA, and PSR) and novel methods such as CRISPR-Cas and microfluidics platforms. Successful molecular diagnostic developments are presented by highlighting their most important findings. Finally, we describe the barriers that hinder the large-scale development of affordable, accessible, rapid, and easy-to-use molecular diagnostic tests. The evolution of diagnostic techniques is critical to prevent the spread of viruses and the development of viral reservoirs in the swine industry that impact the possible development of future pandemics and the world economy.

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Development of a Cleaved Probe-Based Loop-Mediated Isothermal Amplification Assay for Rapid Detection of African Swine Fever Virus

Cited by 4 publications

References 23 publications

Current detection methods of African swine fever virus

Current detection methods of African swine fever virus

Point-of-Care Testing for Hepatitis Viruses: A Growing Need

Emergent Molecular Techniques Applied to the Detection of Porcine Viruses

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