PMID- 36157191 OWN - NLM STAT- PubMed-not-MEDLINE LR - 20220928 IS - 2297-1769 (Print) IS - 2297-1769 (Electronic) IS - 2297-1769 (Linking) VI - 9 DP - 2022 TI - Development of an indirect ELISA for the identification of African swine fever virus wild-type strains and CD2v-deleted strains. PG - 1006895 LID - 10.3389/fvets.2022.1006895 [doi] LID - 1006895 AB - African swine fever (ASF) is a potent infectious disease with detrimental effects on the global swine industry and no currently vaccine available. The emergence of low-virulence CD2v-deleted mutants manifested as non-hemadsorption (non-HAD) strains represents a significant challenge to the prevention and control of ASF. In this study, we aimed to establish an indirect ELISA (IELISA) method for the identification of ASFV wild-type and CD2v-deleted strains. We integrated the CD2v protein extracellular domain sequence (CD2v-Ex, 1-588 bp) of the highly pathogenic strain China/2018/AnhuiXCGQ into the genome of suspension culture-adapted Chinese hamster Ovary-S (CHO-S) cells using lentivirus vectors (LVs). By screening, we identified a monoclonal CHO-S cell line that stably expressed secretory CD2v-Ex Protein. We then used the purified CD2v-Ex Protein as the detection antigen to establish an indirect ELISA method (CD2v-IELISA) for identification of the ASFV wild-type and CD2v-Deleted (CD2v(-)) strains. The CD2v-IELISA method showed excellent specificity with no cross-reaction with serum samples infected with ASFV (CD2v(-)), porcine reproductive and respiratory syndrome virus (PRRSV), classical swine fever virus (CSFV), porcine circovirus (PCV), porcine pseudorabies virus (PRV), swine foot and mouth disease virus (FMDV) and porcine epidemic diarrhea virus (PEDV). Furthermore, this method showed high sensitivity, allowing identification of ASFV-infected clinical serum samples up to a dilution of 1:2,560. The coefficient of variation both in and between batches was <10% with good reproducibility and a high compliance rate of 99.4%. This CD2v-IELISA method developed here is of great significance for the prevention, control and purification of ASFV. CI - Copyright (c) 2022 Jiang, Jiang, Li, Wan, Wang, Wang, Shi, Zhao, Song, Zhu, Ji and Zhang. FAU - Jiang, Wenting AU - Jiang W AD - College of Veterinary Medicine, Henan Agricultural University, Zhengzhou, China. AD - International Joint Research Center of National Animal Immunology, College of Veterinary Medicine, Henan Agricultural University, Zhengzhou, China. FAU - Jiang, Dawei AU - Jiang D AD - College of Veterinary Medicine, Henan Agricultural University, Zhengzhou, China. AD - International Joint Research Center of National Animal Immunology, College of Veterinary Medicine, Henan Agricultural University, Zhengzhou, China. AD - Longhu Laboratory, Zhengzhou, China. FAU - Li, Lu AU - Li L AD - College of Veterinary Medicine, Henan Agricultural University, Zhengzhou, China. AD - International Joint Research Center of National Animal Immunology, College of Veterinary Medicine, Henan Agricultural University, Zhengzhou, China. FAU - Wan, Bo AU - Wan B AD - College of Veterinary Medicine, Henan Agricultural University, Zhengzhou, China. AD - International Joint Research Center of National Animal Immunology, College of Veterinary Medicine, Henan Agricultural University, Zhengzhou, China. AD - Henan Engineering Laboratory of Animal Biological Products, College of Veterinary Medicine, Henan Agricultural University, Zhengzhou, China. FAU - Wang, Jiabin AU - Wang J AD - College of Veterinary Medicine, Henan Agricultural University, Zhengzhou, China. AD - International Joint Research Center of National Animal Immunology, College of Veterinary Medicine, Henan Agricultural University, Zhengzhou, China. FAU - Wang, Panpan AU - Wang P AD - College of Veterinary Medicine, Henan Agricultural University, Zhengzhou, China. AD - International Joint Research Center of National Animal Immunology, College of Veterinary Medicine, Henan Agricultural University, Zhengzhou, China. FAU - Shi, Xuejian AU - Shi X AD - College of Veterinary Medicine, Henan Agricultural University, Zhengzhou, China. AD - International Joint Research Center of National Animal Immunology, College of Veterinary Medicine, Henan Agricultural University, Zhengzhou, China. FAU - Zhao, Qi AU - Zhao Q AD - College of Veterinary Medicine, Henan Agricultural University, Zhengzhou, China. AD - International Joint Research Center of National Animal Immunology, College of Veterinary Medicine, Henan Agricultural University, Zhengzhou, China. FAU - Song, Jinxing AU - Song J AD - College of Veterinary Medicine, Henan Agricultural University, Zhengzhou, China. AD - International Joint Research Center of National Animal Immunology, College of Veterinary Medicine, Henan Agricultural University, Zhengzhou, China. FAU - Zhu, Zixiang AU - Zhu Z AD - State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, College of Veterinary Medicine, Lanzhou University, Chinese Academy of Agricultural Sciences, Lanzhou, China. AD - African Swine Fever Regional Laboratory of China, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, China. FAU - Ji, Pengchao AU - Ji P AD - College of Veterinary Medicine, Henan Agricultural University, Zhengzhou, China. AD - International Joint Research Center of National Animal Immunology, College of Veterinary Medicine, Henan Agricultural University, Zhengzhou, China. AD - Henan Engineering Laboratory of Animal Biological Products, College of Veterinary Medicine, Henan Agricultural University, Zhengzhou, China. FAU - Zhang, Gaiping AU - Zhang G AD - College of Veterinary Medicine, Henan Agricultural University, Zhengzhou, China. AD - International Joint Research Center of National Animal Immunology, College of Veterinary Medicine, Henan Agricultural University, Zhengzhou, China. AD - Longhu Laboratory, Zhengzhou, China. AD - Henan Engineering Laboratory of Animal Biological Products, College of Veterinary Medicine, Henan Agricultural University, Zhengzhou, China. LA - eng PT - Journal Article DEP - 20220908 PL - Switzerland TA - Front Vet Sci JT - Frontiers in veterinary science JID - 101666658 PMC - PMC9493115 OTO - NOTNLM OT - ASFV OT - CD2v extracellular fragment OT - CD2v-deleted OT - CHO cell line OT - identification OT - indirect ELISA COIS- The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. EDAT- 2022/09/27 06:00 MHDA- 2022/09/27 06:01 PMCR- 2022/01/01 CRDT- 2022/09/26 17:04 PHST- 2022/07/29 00:00 [received] PHST- 2022/08/22 00:00 [accepted] PHST- 2022/09/26 17:04 [entrez] PHST- 2022/09/27 06:00 [pubmed] PHST- 2022/09/27 06:01 [medline] PHST- 2022/01/01 00:00 [pmc-release] AID - 10.3389/fvets.2022.1006895 [doi] PST - epublish SO - Front Vet Sci. 2022 Sep 8;9:1006895. doi: 10.3389/fvets.2022.1006895. eCollection 2022.