Establishment of an indirect immunofluorescence assay for the detection of African swine fever virus antibodies

作     者：Wan Wang Zhenjiang Zhang Weldu Tesfagaber Jiwen Zhang Fang Li Encheng Sun Lijie Tang Zhigao Bu Yuanmao Zhu Dongming Zhao Wan Wang;Zhenjiang Zhang;Weldu Tesfagaber;Jiwen Zhang;Fang Li;Encheng Sun;Lijie Tang;Zhigao Bu;Yuanmao Zhu;Dongming Zhao

作者机构：State Key Laboratory for Animal Disease Control and PreventionNational African Swine Fever Para-reference LaboratoryNational High Containment Facilities for Animal Diseases Control and PreventionHarbin Veterinary Research InstituteChinese Academy of Agricultural SciencesHarbin 150069China College of Veterinary MedicineNortheast Agricultural UniversityHarbin 150069China 

出 版 物：《Journal of Integrative Agriculture》 (农业科学学报（英文版）)

年 卷 期：2024年第23卷第1期

页      面：228-238页

核心收录：

学科分类：090602[农学-预防兽医学] 09[农学] 0906[农学-兽医学] 0901[农学-作物学] 

基　　金：supported by the National Key R&D Program of China(2019YFE0107300 and 2021YFD1800101) the Applied Technology Research and Development Project of Heilongjiang Province,China(GA19B301) the Central Public-interest Scientific Institution Basal Research Fund,China(1610302022003) 

主　　题：African swine fever antibody IFA serological method 

摘      要：African swine fever(ASF)continues to cause enormous economic loss to the global pig *** there is no safe and effective vaccine,accurate and timely diagnosis of ASF is essential to implement control *** 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 *** 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 *** IFA reacted with the ASFV-positive sera and displayed bright fluorescence *** was no non-specific green fluorescence due to cellular senescence or other cell damage-causing *** to a commercial indirect enzyme-linked immunosorbent assay(iELISA),ASFV antibodies were detected 1–4 days earlier using our *** detection limits of the IFA and iELISA for the same ASFV-antibody positive serum samples were 1:25,600 and 1:6,400,respectively,indicating that the IFA is more sensitive than *** 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 type O(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.