Real-Time Detection of LAMP Products of African Swine Fever Virus Using Fluorescence and Surface Plasmon Resonance Method

Abstract

African swine fever (ASF) is a swine disease with a very high fatality rate caused by a complex double-stranded DNA virus. The fluorescence PCR detection method is widely used for virus nucleic acid detection. Surface plasmon resonance (SPR) is a label-free and real-time detection method, unlike the fluorescence PCR detection method. In this research, we detected the loop-mediated isothermal amplification (LAMP) products of the African swine fever virus by using the SPR and fluorescence methods separately and simultaneously. By comparing the positive and negative control results, we found that the SPR response unit is completely different before and after the LAMP process. In addition, the fluorescence results on a chip showed that with an increase in the concentration of the sample, the cycle threshold (CT) value decreased, which is consistent with commercial instruments. Both the decline rate of the SPR response unit and the CT value of the fluorescence realized were used to distinguish the positive control from the negative control and water, which indicates that the SPR method can be combined with fluorescence to detect LAMP products. This research provides a label-free and simple method for detecting LAMP products.

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“…This method was supported by the provided evidence that non-water-borne viruses can be detected during wastewater monitoring. For example, airborne viruses (coronavirus [121] and influenza [133]) and vector-borne viruses (Zika virus [134] and West Nile virus [135]) were detected via wastewater-based epidemiological studies.…”

“…In addition, if the target sequence mutates significantly, the PCR may not be able to detect the new strains that diverged from the target virus. Other than PCR, other virus detection methods include virus culture [120], loopmediated isothermal amplification (LAMP) [121], enzyme-linked immunosorbent assay (ELISA) [122], immunofluorescence assay (IFA) [123], and next-generation sequencing (NGS) [124]. Due to the ability to sequence genetic material from samples, NGS is widely used to identify unknown viruses [125].…”