Abstract
Porcine epidemic diarrhea virus (PEDV), as the main causative pathogen of viral diarrhea in pigs, has been reported to result in high morbidity and mortality in neonatal piglets and cause significant economic losses to the swine industry. Rapid diagnosis methods are essential for preventing outbreaks and transmission of this disease. In this study, a paper-based lateral flow immunoassay for the rapid diagnosis of PEDV in swine fecal samples was developed using stable color-rich latex beads as the label. Under optimal conditions, the newly developed latex bead-based lateral flow immunoassay (LBs-LFIA) attained a limit of detection (LOD) as low as 103.60 TCID50/mL and no cross-reactivity with other related swine viruses. To solve swine feces impurity interference, by adding a filtration unit design of LFIA without an additional pretreatment procedure, the LBs-LFIA gave good agreement (92.59%) with RT-PCR results in the analysis of clinical swine fecal samples (n = 108), which was more accurate than previously reported colloidal gold LFIA (74.07%) and fluorescent LFIA (86.67%). Moreover, LBs-LFIA showed sufficient accuracy (coefficient of variance [CV] < 15%) and stable (room temperature storage life > 56 days) performance for PEDV detection, which is promising for on-site analysis and user-driven testing in pig production system.
Highlights
The rapid diagnosis of pathogenic microorganisms is essential to identify diseases and provide the correct preventive medicine or treatment (Carter et al 2020; Wang et al 2020a)
Principle of the latex beads (LBs)-LFIA for Porcine epidemic diarrhea virus (PEDV) detection Once the viral analyte reaches the conjugate pad, it is recognized by specific detection antibodies, and the immune complexes continue to move along until they are captured at the test line via a predeposited capturing antibody, which forms a double-antibody sandwich structure
The aggregated latex beads on test line (T line) are dependent on the presence and concentration of PEDV; their concentration can be measured by a colorimetric assay
Summary
The rapid diagnosis of pathogenic microorganisms is essential to identify diseases and provide the correct preventive medicine or treatment (Carter et al 2020; Wang et al 2020a). The predominant trend is the development of pragmatic means to monitor animal health, with a greater focus on preventive medicine rather than treatment after a disease is contracted. Porcine epidemic diarrhea virus (PEDV), of the genus Alphacoronavirus in the family Coronaviridae, causes watery diarrhea, vomiting and dehydration, and has. As an acute and highly contagious enteric disease, porcine epidemic diarrhea (PED) caused by PEDV was first reported in England and Belgium, and the emergence and reemergence of PED outbreaks have occurred in Europe, America and Asia in recent decades (Diep et al 2018; Pensaert and de Bouck 1978; Sun et al 2012; Wood 1977). The threat posed by PEDV still persists (Chen et al 2019; Gao et al 2013)
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