PMID- 36253821 OWN - NLM STAT- MEDLINE DCOM- 20221019 LR - 20221021 IS - 1746-6148 (Electronic) IS - 1746-6148 (Linking) VI - 18 IP - 1 DP - 2022 Oct 18 TI - Development of gold Immunochromatographic assay strip based on specific polyclonal antibodies against capsid protein for rapid detection of porcine circovirus 2 in Zhejiang province, China. PG - 373 LID - 10.1186/s12917-022-03471-6 [doi] LID - 373 AB - BACKGROUND: The existing detection methods for porcine circovirus type 2 (PCV2) specific antibodies in serum cannot determine the infection status, thus it is necessary to establish a method for detecting PCV2 antigen. The capsid protein (CAP) of PCV2, as a major structural protein that plays a significant role in viral replication and in inducing host's immune response, is an ideal target antigen to monitor PCV2 infection. Therefore, a gold immunochromatographic assay (GICA) for rapid detection of PCV2 antigen based on the polyclonal antibodies (PAbs) against PCV2-CAP will be developed. RESULTS: The truncated CAP protein (dCAP) was used to immunize rabbits to generate anti-serum. After preliminary purification by caprylic acid/ammonium sulfate precipitation (CAAS), specific PAbs were purified by affinity chromatography column coupled with dCAP and its titer was about two-fold higher than preliminary purified PAbs. Colloidal gold-PAbs conjugate was synthesized under the optimum conditions. The specific anti-dCAP PAbs and goat anti-rabbit antibody (GAR) were then sprayed onto nitrocellulose (NC) membrane as a test line (TL) and a control line (CL), respectively. The visual limit detection (vLOD) of the GICA strips was 5 ng/mL. Specificity assay indicated that the GICA strips had specifically detected PCV2 and was not reactive for porcine epidemic diarrhea virus (PEDV), pseudorabies virus (PRV), porcine reproductive and respiratory syndrome virus (PRRSV) or classic swine fever virus (CSFV). A total of 36 porcine serum samples were detected by this GICA and commercial enzyme-linked immunosorbent assay (ELISA) Kit, 9 positive samples were found by the developed strip with the rate of 25.0% comparing with 11 positive samples detected by the commercially ELISA Kit which positive rate was 30.5%, and the receiver operating characteristic (ROC) curve revealed that the relative sensitivity and specificity of this GICA strip were 72.7 and 96.0%, respectively, with an area of 87.2%. CONCLUSIONS: This study established an efficient detection method with high sensitivity and specificity for the clinical diagnosis of PCV2 antigen, that will facilitate a rapid and convenient way to evaluate the infection status of vaccinated pigs. CI - (c) 2022. The Author(s). FAU - Ding, Haojie AU - Ding H AD - School of Basic Medical Sciences and Forensic Medicine, Hangzhou Medical College, Hangzhou, Zhejiang, China. FAU - Shen, Yu AU - Shen Y AD - School of Basic Medical Sciences and Forensic Medicine, Hangzhou Medical College, Hangzhou, Zhejiang, China. FAU - Gao, Yafan AU - Gao Y AD - School of Basic Medical Sciences and Forensic Medicine, Hangzhou Medical College, Hangzhou, Zhejiang, China. FAU - Wu, Songrui AU - Wu S AD - School of Basic Medical Sciences and Forensic Medicine, Hangzhou Medical College, Hangzhou, Zhejiang, China. FAU - Xie, ChengZuo AU - Xie C AD - School of Basic Medical Sciences and Forensic Medicine, Hangzhou Medical College, Hangzhou, Zhejiang, China. FAU - Sun, Hao AU - Sun H AD - School of Basic Medical Sciences and Forensic Medicine, Hangzhou Medical College, Hangzhou, Zhejiang, China. FAU - Zhang, Hongli AU - Zhang H AD - Zhejiang Center of Animal Disease Control, Hangzhou, Zhejiang, China. FAU - Sun, Hongchao AU - Sun H AD - Department of Animal Parasitology, Institute of Animal Husbandry and Veterinary Medicine, Zhejiang Academy of Agricultural Science, Hangzhou, Zhejiang, China. FAU - Shan, Ying AU - Shan Y AD - Department of Veterinary Medicine, College of Animal Sciences, Zhejiang University, Hangzhou, Zhejiang, China. FAU - Ding, Jianzu AU - Ding J AD - School of Basic Medical Sciences and Forensic Medicine, Hangzhou Medical College, Hangzhou, Zhejiang, China. FAU - Zheng, Bin AU - Zheng B AD - School of Basic Medical Sciences and Forensic Medicine, Hangzhou Medical College, Hangzhou, Zhejiang, China. FAU - Lu, Shaohong AU - Lu S AD - School of Basic Medical Sciences and Forensic Medicine, Hangzhou Medical College, Hangzhou, Zhejiang, China. llsshh2003@163.com.cn. FAU - Zhuo, Xunhui AU - Zhuo X AD - School of Basic Medical Sciences and Forensic Medicine, Hangzhou Medical College, Hangzhou, Zhejiang, China. Zhuoxh@zjams.com.cn. LA - eng PT - Journal Article DEP - 20221018 PL - England TA - BMC Vet Res JT - BMC veterinary research JID - 101249759 RN - 0 (Antibodies, Viral) RN - 0 (Capsid Proteins) RN - 0 (Gold Colloid) RN - 9004-70-0 (Collodion) RN - SU46BAM238 (Ammonium Sulfate) SB - IM MH - Ammonium Sulfate MH - Animals MH - Antibodies, Viral MH - Capsid Proteins MH - *Circoviridae Infections/diagnosis/veterinary MH - *Circovirus MH - Collodion MH - Gold Colloid/chemistry MH - Immunoassay/veterinary MH - Rabbits MH - Swine MH - *Swine Diseases PMC - PMC9578217 OTO - NOTNLM OT - Capsid protein OT - Gold immunochromatographic assay OT - Polyclonal antibody OT - Porcine circovirus type 2 COIS- The authors declare that they have no competing interests. EDAT- 2022/10/18 06:00 MHDA- 2022/10/20 06:00 PMCR- 2022/10/18 CRDT- 2022/10/17 23:46 PHST- 2022/05/17 00:00 [received] PHST- 2022/10/04 00:00 [accepted] PHST- 2022/10/17 23:46 [entrez] PHST- 2022/10/18 06:00 [pubmed] PHST- 2022/10/20 06:00 [medline] PHST- 2022/10/18 00:00 [pmc-release] AID - 10.1186/s12917-022-03471-6 [pii] AID - 3471 [pii] AID - 10.1186/s12917-022-03471-6 [doi] PST - epublish SO - BMC Vet Res. 2022 Oct 18;18(1):373. doi: 10.1186/s12917-022-03471-6.