Key result
A LAMP-CRISPR assay detected the ASFV p72 gene with a limit of 7 copies/μl per reaction, showing no cross-reactivity and good consistency with real-time qPCR for clinical samples.
Why the study?
African swine fever is one of the most severe infectious diseases of pigs, necessitating convenient, portable, low-cost, sensitive, and specific detection methods.
Does a LAMP-CRISPR assay accurately and sensitively detect African swine fever virus compared to real-time qPCR?
Does a LAMP-CRISPR assay accurately and sensitively detect African swine fever virus compared to real-time qPCR?
The LAMP-CRISPR assay provides a highly sensitive, specific, and portable method for on-site detection of African swine fever virus.
One-tube LAMP-CRISPR assay enables rapid ASFV detection; leaves open field validation before routine diagnostic use.
African swine fever (ASF) is one of the most severe infectious diseases of pigs. In this study, a loop-mediated isothermal amplification (LAMP) assay coupled with the clustered regularly interspaced short palindromic repeats (CRISPR)/Cas12a system was established in one tube for the detection of the African swine fever virus (ASFV) p72 gene. The single-stranded DNA-fluorophore quencher reporter and CRISPR-derived RNA were screened and selected for the CRISPR detection system. In combination with LAMP amplification assay, the detection limit for the LAMP-CRISPR assay can reach 7 copies/μl of p72 gene per reaction. Furthermore, this method displays no cross-reactivity with other porcine DNA or RNA viruses. The performance of the LAMP-CRISPR assay was compared with real-time qPCR tests for clinical samples; a good consistency between the LAMP-CRISPR assay and real-time qPCR was observed. The method shed a light on the convenient, portable, low cost, highly sensitive and specific detection of ASFV, demonstrating a great application potential for monitoring on-site ASFV in the field.
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Yang et al. (2021) studied African swine fever (ASF). LAMP assay coupled with CRISPR/Cas12a system vs. real-time qPCR was evaluated on Detection limit of ASFV p72 gene. A LAMP-CRISPR assay detected the ASFV p72 gene with a limit of 7 copies/μl per reaction, showing no cross-reactivity and good consistency with real-time qPCR for clinical samples.
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