PCV4 Cap抗体ELISA检测方法的建立及血清流行病学调查

doi:10.11843/j.issn.0366-6964.2024.05.025

Abstract

Abstract: The present study aimed to develop an indirect ELISA to detect antibodies against the Cap protein of porcine circovirus type 4 (PCV4), and to detect PCV4 antibodies in pigs, then compare the results with previous investigations on PCV4 seroepidemiology, so as to understand the prevalence and infection of PCV4 in swine populations more comprehensively. PCV4 Cap protein was successfully expressed in a prokaryotic expression system. By optimizing the reaction conditions, an indirect ELISA method based on PCV4 Cap protein was established. To investigate the serological prevalence of PCV4 in Chinese pig herds, 2 298 serum samples from pigs at different growth stages from 18 provinces in China were analyzed using the established ELISA method. Concurrently, the Cap-ELISA method was compared with Rep-ELISA, and 845 serum samples were tested in parallel. The results showed that PCV4 antibodies were detected in serum samples from 17 out of the 18 provinces. The total seroprevalence of PCV4 was 34.94%, and the positive rates of sows and finishing pigs were 59.95% and 31.64%, respectively. The positive rate of piglets was 21.14%, while nursery pigs had the lowest positive rate at 4.20%. A total of 845 serum samples were tested using both methods, and the coincidence rate was 82.84%. The Cap-ELISA test yielded an overall positive rate of 31.83%, while the Rep-ELISA test had an overall positive rate of 35.03%. Studies have demonstrated the widespread prevalence of PCV4 across the country and its ability to infect pigs of all ages. The Cap-ELISA and Rep-ELISA techniques exhibited a high degree of concordance, and based on the antibody growth and decline trends, both methods indicated that PCV4 infection primarily occurs in finishing pigs and sows. The present study provides the most recent data on the seroepidemiological characteristics of PCV4 in China and PCV4 infection prevalence in pigs at different growth stages, which could facilitate further research on PCV4 and its management.

Key words: PCV4, prokaryotic expression, indirect ELISA, seroepidemiological survey

CLC Number:

S852.659.2

SONG Xiaoqing, DENG Ruide, LI Xin, LI Jiao, LI Runcheng, DU Lifei, DONG Wei, GE Meng. Establishment of ELISA for Detection of PCV4-Cap Antibody and Sero-epidemiological Survey[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(5): 2072-2079.

References

[1] ZHANG H H, HU W Q, LI J Y, et al. Novel circovirus species identified in farmed pigs designated as Porcine circovirus 4, Hunan province, China[J]. Transbound Emerg Dis, 2020, 67(3):1057-1061.
[2] TISCHER I, GELDERBLOM H, VETTERMANN W, et al. A very small porcine virus with circular single-stranded DNA[J]. Nature, 1982, 295(5844):64-66.
[3] TISCHER I, MIELDS W, WOLFF D, et al. Studies on epidemiology and pathogenicity of porcine circovirus[J]. Arch Virol, 1986, 91(3-4):271-276.
[4] ELLIS J, HASSARD L, CLARK E, et al. Isolation of circovirus from lesions of pigs with postweaning multisystemic wasting syndrome[J]. Can Vet J, 1998, 39(1):44-51.
[5] SAOULIDIS K, KYRIAKIS S C, KENNEDY S, et al. First report of post-weaning multisystemic wasting syndrome and porcine dermatitis and nephropathy syndrome in pigs in Greece[J]. J Vet Med B, 2002, 49(4):202-205.
[6] JIANG H J, WANG D, WANG J, et al. Induction of porcine dermatitis and nephropathy syndrome in piglets by infection with porcine circovirus type 3[J]. J Virol, 2019, 93(4):e02045-18.
[7] PALINSKI R, PIÑEYRO P, SHANG P C, et al. A novel porcine circovirus distantly related to known circoviruses is associated with porcine dermatitis and nephropathy syndrome and reproductive failure[J]. J Virol, 2016, 91(1):e01879-16.
[8] 唐宁, 吕艳丽, 郭鑫, 等. 免疫组化法检测猪圆环病毒2型在人工感染猪体内的分布[J]. 畜牧兽医学报, 2008, 39(3):372-375.
TANG N, LÜ Y L, GUO X, et al. Detection of distribution of porcine circovirus type 2 in experimentally infected piglets with immunohistochemical method[J]. Acta Veterinaria et Zootechnica Sinica, 2008, 39(3):372-375. (in Chinese)
[9] 库旭钢, 于学祥, 孙琪, 等. 猪圆环病毒3型的分离及致病性分析[J]. 畜牧兽医学报, 2023, 54(4):1568-1578.
KU X G, YU X X, SUN Q, et al. Isolation and pathogenicity analysis of porcine circovirus type 3[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(4):1568-1578. (in Chinese)
[10] NGUYEN V G, DO H Q, HUYNH T M L, et al. Molecular-based detection, genetic characterization and phylogenetic analysis of porcine circovirus 4 from Korean domestic swine farms[J]. Transbound Emerg Dis, 2022, 69(2):538-548.
[11] SIRISEREEWAN C, NGUYEN T C, PIEWBANG C, et al. Molecular detection and genetic characterization of porcine circovirus 4 (PCV4) in Thailand during 2019-2020[J]. Sci Rep, 2023, 13(1):5168.
[12] XU T, CHEN L, HUANG B Z, et al. The first dog-origin porcine circovirus type 4 complete genomic sequence have high homology with that of pig-derived strains[J]. Front Microbiol, 2023, 14:1121177.
[13] ZHANG L H, WANG T X, FU P F, et al. First molecular detection and genetic analysis of a novel porcine circovirus (porcine circovirus 4) in dogs in the world[J]. Microbiol Spectr, 2023, 11(2):e0433322.
[14] XU T, CHEN X M, FU Y, et al. Cross-species transmission of an emerging porcine circovirus (PCV4):first molecular detection and retrospective investigation in dairy cows[J]. Vet Microbiol, 2022, 273:109528.
[15] WANG Y J, YAN S J, JI Y T, et al. First identification and phylogenetic analysis of porcine circovirus type 4 in fur animals in Hebei, China[J]. Animals, 2022, 12(23):3325.
[16] HOU C Y, ZHANG L H, ZHANG Y H, et al. Phylogenetic analysis of porcine circovirus 4 in Henan Province of China:a retrospective study from 2011 to 2021[J]. Transbound Emerg Dis, 2022, 69(4):1890-1901.
[17] SUN W C, DU Q, HAN Z X, et al. Detection and genetic characterization of porcine circovirus 4 (PCV4) in Guangxi, China[J]. Gene, 2021, 773:145384.
[18] TIAN R B, ZHAO Y, CUI J T, et al. Molecular detection and phylogenetic analysis of Porcine circovirus 4 in Henan and Shanxi Provinces of China[J]. Transbound Emerg Dis, 2021, 68(2):276-282.
[19] NIU G Y, ZHANG X W, JI W L, et al. Porcine circovirus 4 rescued from an infectious clone is replicable and pathogenic in vivo[J]. Transbound Emerg Dis, 2022, 69(5):e1632-e1641.
[20] GE M, HU W Q, NING K M, et al. The seroprevalence of the newly identified porcine circovirus type 4 in China investigated by an enzymed-linked immunosorbent assay[J]. Transbound Emerg Dis, 2021, 68(6):2910-2914.
[21] LIAN Z M, LIU J B, LIU P R, et al. Development and application of an indirect ELISA for the detection of antibody to porcine circovirus 4 in pigs[J]. Transbound Emerg Dis, 2021, 68(6):2975-2979.
[22] HU X F, DING Z, LI Y, et al. Serum investigation of antibodies against porcine circovirus 4 Rep and Cap protein in Jiangxi Province, China[J]. Front Microbiol, 2022, 13:944679.

Establishment of ELISA for Detection of PCV4-Cap Antibody and Sero-epidemiological Survey

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	HUANG Jin, LI Siyuan, MAO Li, CAI Xuhang, XIE Lingling, WANG Fu, ZHOU Hua, LI Jizong, LI Bin. Eukaryotic Expression of Bovine Coronavirus S1 Protein and Establishment and Application of Indirect ELISA [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(5): 2050-2060.
[2]	ZHAO Canqi, FENG Yu, LÜ Lang, LI Yanjun, WEI Yulei, DING Jiabo, CHEN Xiang, JIANG Hui. Study on Purification of Bovine Brucellosis by Competitive ELISA and Indirect ELISA [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(5): 2146-2153.
[3]	LUO Xiaofen, XIE Xiaodong, ZHAO Chao, HU Qian, WANG Yongxuan, RAN Fangfei, HU Pengfei, WEN Ming, ZHU Erpeng, CHENG Zhentao. Initial Identification of Adhesion-related Proteins of Mycoplasma bovis of Guizhou Strains [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(4): 1672-1683.
[4]	LIU Chuanxia, WANG Xiao, LI Xuewen, BAO Miaofei, LI Tingting, CHEN Xin, WENG Changjiang, ZHENG Jun. Preparation of Monoclonal Antibody of African Swine Fever Virus pE120R [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(1): 388-394.
[5]	WANG Jingyu, PAN Yangyang, XU Gengquan, ZHANG Rui, ZHANG Wenlan, WANG Xiaoshan, WU Rentaodi, ZHAO Rigetu, CUI Yan, YU Sijiu. Preparation and Preliminary Application of Yak (Bos Grunniens) Fas-associated Factor 1 Polyclonal Antibody [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(8): 3369-3382.
[6]	LIU Taoxue, SU Bingqian, QI Yanli, GUO Jiangtao, LIU Zhonghu, CHU Beibei, WANG Jiang, ZENG Lei. Preparation of the Monoclonal Antibody against the African Swine Fever Virus p30 Protein and Identification of the Antigenic Epitope [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(8): 3415-3423.
[7]	JING Tingting, YIN Hao, HUANG Rong, LAN Shimei, LI Zhangcheng, YOU Liuchao, HAO Huafang, FU Lei, CHU Yuefeng. Prokaryotic Expression and Biological Function Analysis of Mycoplasma bovis 0580 Gene C-terminal Truncated [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(7): 2991-3001.
[8]	WANG Hui, XU Mengyuan, LIU Lingkang, ZHENG Xibang, LI Gonghe, WU Wende. PhiC31 Integrase Triggers Site-specific Cassette Exchange in Chicken Fibroblast Cells through Electroporation [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(6): 2330-2342.
[9]	JIAO Zhengxing, PAN Yangyang, WANG Meng, WANG Jinglei, MA Wenbin, GAO Xiang, ZHANG Hui, CUI Yan, YU Sijiu, WANG Libin. Preparation of Polyclonal Antibody against Yak LC3B Protein and Its Application in Detection of Expression in Reproductive Organs [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(6): 2436-2447.
[10]	GAO Zhenzhen, MENG Zejing, HE Xiaobing, FANG Yongxiang, TIAN Huihui, CHEN Guohua, JING Zhizhong. Identification of Immunogenicity and Neutralizing Activity of Ectromelia Virus EVM135 and EVM085 Proteins [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(6): 2468-2477.
[11]	CHENG Tianyin, WU Congying, LIU Yuke, DUAN Deyong. Studies on the Role of HSP70-b2 and Its 14-Mer-like Peptide from Haemaphysalis flava [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(6): 2570-2580.
[12]	WANG Guochao, ZHAO Yaru, ZHANG Zhonghui, ZHANG Yulong, BAI Ge, GENG Shuxian, FAN Jie, YANG Jifei, GUAN Guiquan, YIN Hong, LUO Jianxun, NIU Qingli. Bioinformatics Analysis of RNA Polymerase Subunit D205R Gene of African Swine Fever Virus and Polyclonal Antibody Preparation [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(5): 2042-2049.
[13]	ZHAI Yunyi, YUAN Ye, LI Junmei, TIAN Lulu, DIAO Ziyang, LI Bin, CHEN Jialu, ZHOU Dong, JIN Yaping, WANG Aihua. Preparation and Preliminary Application of Monoclonal Antibody to Brucella Outer Membrane Protein 16 [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(5): 2083-2091.
[14]	WANG Bingnan, SUN Xue, JIA Hongguo, TAN Chun, MEI Sipeng, ZHOU Rongqiong. Prokaryotic Expression and Tissue Localization of C-type Lectin 4 from Toxocara canis [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(4): 1641-1651.
[15]	CAI Mingyu, ZHANG Hailong, HAI Zhenzhen, QIAO Yarui, DU Jun, ZHOU Xuezhang. The Inflamed Molecular Mechanism Induced by Recombined 14-3-3 Protein of Candida krusei on Bovine Mammary Epithelial Cells [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(4): 1679-1689.