Development of a duplex lateral flow assay for simultaneous detection of antibodies against African and Classical swine fever viruses

Abstract

Classical swine fever (CSF) and African swine fever (ASF) are both highly contagious diseases of domestic pigs and wild boar and are clinically indistinguishable. For both diseases, antibody detection is an integral and crucial part of prevention and control measures. The purpose of our study was to develop and initially validate a duplex pen-side test for simultaneous detection and differentiation of specific antibodies against CSF virus (CSFV) and ASF virus (ASFV). The test was based on the major capsid protein VP72 of ASFV and the structural protein E2 of CSFV, both considered the most immunogenic proteins of these viruses. The performance of the pen-side test was evaluated using a panel of porcine samples consisting of experimental, reference, and field sera, with the latter collected from European farms free of both diseases. The new lateral flow assay was able to detect specific antibodies to ASFV or CSFV, showing good levels of sensitivity and specificity. These preliminary data indicate the potential of the newly developed pen-side test for rapid differential detection of antibodies found in the 2 diseases, which is of particular importance in the field and in front-line laboratories where equipment and skilled personnel are limited and control of ASF and CSF is crucial.

Characterization of the highly immunogenic VP2 protrusion domain as a diagnostic antigen for members of Birnaviridae family

“…S6). Capsid protein-based lateral flow assays have recently received increased attention due to their ease-of-use and cost-effectiveness (Sun et al 2014;Sastre et al 2016). A capsid protein-based lateral flow assay is currently not available for birnaviruses.…”

“…Recently, lateral flow assays, also known as strip-based immunoassays, which are based on capsid proteins have gained considerable attention for detection of various pathogens since these assays are rapid, user-friendly, and costeffective (Sun et al 2014;Sastre et al 2016). Moreover, technical expertise or special instruments are not required for performing lateral flow assays.…”

Gaps in African swine fever: Analysis and priorities

“…New technologies including lateral flow devices (pen‐side tests) and portable PCR machines that allow rapid diagnosis have been recently developed (Sastre, Gallardo, et al., ; Sastre, Pérez, et al., ). A deeper validation under field conditions should be encouraged.…”

“…and portable PCR machines that allow rapid diagnosis have been recently developed (Sastre, Gallardo, et al, 2016;Sastre, P erez, et al, 2016). 2015).…”

Multiplex Lateral Flow Immunoassay: An Overview of Strategies Towards High-Throughput Point-of-Need Testing

“…Considered that LFDs are intended for being operated by untrained personnel, often multicolored labels have been used to facilitate distinguishing the various detected analytes. In particular, spatially separated detection sites were identified not only based on positioning of the line along the strip, but also on the specific color of the probe used [34,[42][43][44].…”