Key result
The newly established indirect immunofluorescence assay detected ASFV antibodies 1-4 days earlier and was more sensitive (detection limit 1:25600 vs 1:6400) than a commercial iELISA.
Why the study?
Because there is no safe and effective vaccine for African swine fever, accurate and timely diagnosis is essential to implement control measures.
The newly established IFA using a wild boar cell line provides a highly sensitive and specific method for the serological diagnosis of African swine fever virus.
May enable earlier ASFV serodiagnosis in swine; leaves open prospective validation before routine use.
African swine fever (ASF) continues to cause enormous economic loss to the global pig industry. Since there is no safe and effective vaccine, accurate and timely diagnosis of ASF is essential to implement control measures. Indirect immunofluorescence assay (IFA) is a gold standard serological method recommended by the World Organization for Animal Health (WOAH). In this study, we used primary fetal kidney cells to establish a wild boar cell line (BK2258) that supported the efficient replication of ASF virus (ASFV) SD/DY-I/21 and showed visible cytopathic effect (CPE). Moreover, using BK2258, we established a sensitive and specific IFA for ASFV antibody detection. To standardize and evaluate the performance of this assay, we used serum samples from pigs infected with the low virulent genotype I SD/DY-I/21 and genotype II HLJ/HRB1/20, and immunized with the vaccine candidate HLJ/18-7GD, field samples, and negative serum samples. The IFA reacted with the ASFV-positive sera and displayed bright fluorescence foci. There was no non-specific green fluorescence due to cellular senescence or other cell damage-causing factors. Compared to a commercial indirect Enzyme-Linked Immunosorbent Assay (iELISA), ASFV antibodies were detected 1-4 days earlier using our IFA. The detection limits of the IFA and iELISA for the same ASFV-antibody positive serum samples were 1:25600 and 1:6400, respectively, indicating that the IFA is more sensitive than iELISA. The newly established IFA was highly specific and did not cross-react with sera positive for six other important porcine pathogens (i.e., Classical swine fever virus (CSFV), Porcine reproductive and respiratory syndrome virus (PRRSV), Porcme circovirus type 2 (PCV2), Pseudorabies virus (PRV), Foot-and-Mouth disease virus (FMDV/O), and Porcine epidemic diarrhea virus (PEDV)). This study thus provides a sensitive, specific, and reliable detection method that is suitable for the serological diagnosis of ASF.
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Wang et al. (2023) studied African swine fever (ASF). Indirect immunofluorescence assay (IFA) using BK2258 cell line vs. Commercial indirect Enzyme-Linked Immunosorbent Assay (iELISA) was evaluated on Detection of ASFV antibodies. The newly established indirect immunofluorescence assay detected ASFV antibodies 1-4 days earlier and was more sensitive (detection limit 1:25600 vs 1:6400) than a commercial iELISA.
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