Abstract
Cross-border pathogens such as the African swine fever virus (ASFV) still pose a socio-economic threat. Cheaper, faster, and accurate diagnostics are imperative for healthcare and food safety applications. Currently, the discovery of the Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) has paved the way for the diagnostics based on Cas13 and Cas12/14 that exhibit collateral cleavage of target and single-stranded DNA (ssDNA) reporter. The reporter is fluorescently labeled to report the presence of a target. These methods are powerful; however, fluorescence-based approaches require expensive apparatuses, complicate results readout, and exhibit high-fluorescence background. Here, we present a new CRISPR–Cas-based approach that combines polymerase chain reaction (PCR) amplification, Cas12a, and a probe-based lateral flow biosensor (LFB) for the simultaneous detection of seven types of ASFV. In the presence of ASFVs, the LFB responded to reporter trans-cleavage by naked eyes and achieved a sensitivity of 2.5 × 10−15 M within 2 h, and unambiguously identified ASFV from swine blood. This system uses less time for PCR pre-amplification and requires cheaper devices; thus, it can be applied to virus monitoring and food samples detection.
Highlights
African swine fever (ASF) is a highly contagious, fatal infectious disease of domestic and wild pigs caused by an Asfarviridae family DNA virus called the African swine fever virus (ASFV) [1].It can be expressed in three main forms: acute, subacute, and chronic [2,3]
ASFV is one of the most detrimental viruses occurring in swine, with high morbidity and ASFV is one of the most detrimental viruses occurring in swine, with high morbidity and mortality, causing severe socio-economic concerns
The early detection of ASFV is crucial for early mortality, causing severe socio-economic concerns
Summary
African swine fever (ASF) is a highly contagious, fatal infectious disease of domestic and wild pigs caused by an Asfarviridae family DNA virus called the African swine fever virus (ASFV) [1]. It can be expressed in three main forms: acute, subacute, and chronic [2,3]. Is stillinto a major in countries worldwide, At present, detection of ASFV beASFV divided twoproblem categories according to different including developing and developed countries such as Russia and countries in Western
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