猪伪狂犬病病毒多克隆抗体的制备及免疫学活性研究

doi:10.16431/j.cnki.1671-7236.2015.06.014

Abstract

Abstract: This study was aimed to obtain polyclonal antibody against swine pseudorabies virus (PRV) Min A strain,and provide a theoretical basis for the study of the treatment and detection of PRV.This study was performed on PK-15 cell and proliferation of PRV was measured as TCID₅₀ 10^-7.372,the protein concentration of PRV was measured as 3.6 mg/mL.Choosing five healthy male rabbits (2.5 kg±0.2 kg) as experimental animals and using PRV obtained as the antigen,we got polyclonal antibody against PRV.Antiserum titer was 1:32 000,antigen coating dilution was 1:40,the best coating conditions was 4 ℃ 12 h,the best blocking time was 1 h,the best working dilution of enzyme labled antibody was 1:8 000,the result of cell lesions neutralization test showed that PRV antiserum prepared in this assay at 1:16 dilution could protect 50% of PK-15 cells from being infected by PRV,and negative serum couldn't protect PK-15 cells from being infected by PRV.The study successfully prepared polyclonal antibodies against PRV.

Key words: pseudorabies virus (PRV); polyclonal antibody; PK-15 cell

CLC Number:

S852.4

MENG Ri-zeng, LIU Tao, MA Wen-chen, WANG Wei-lin, WU Lian-peng, LUO Yan-fei. Preparation of Polyclonal Antibodies against Swine Pseudorabies Virus and Study on the Immunological Activity[J]. , 2015, 42(6): 1417-1423.

References

[1] 童光志,陈焕春.伪狂犬病流行现状及我国应采取的防制措施[J].中国兽医学报,1999,19(1):1-2.

[2] 斯特劳B E.赵德明等译.猪病学[M].北京:中国农业大学出版社,2000.

[3] 孙圣福,陈静,马慧玲,等.不同日龄猪伪狂犬抗体跟踪监测与分析[J].中国畜牧兽医,2011,38(4):45-48.

[4] Halling Shirley M,Peterson-Burch Brooke D,Bricker Betsy J,et al.Completion of the genome sequence of Brucella abortus and comparison to the highly similar genomes of Brucella melitensis and Brucella suis[J]. Journal of Bacteriology,2005,187(8):2715-2726.

[5] 吕超超,焦连国,张金双,等.2010年上半年11省市规模猪场疫病监测与剖析[J].中国畜牧兽医,2011,38(2):25-29.

[6] Satoshi A,Akihiko K,Takahashi T,et al.Monoclonal antibody 3-9-16 recognizes one of the two iosforms of rabies virus matrix protein that exposes its N-terminus on the virion surface[J].Microbio Immnol,2003,47(9):639-651.

[7] 姚敬明,王娟萍,韩一超,等.猪伪狂犬病病毒gE基因缺失苗免疫试验[J].中国畜牧兽医,2011,38(2):56-60.

[8] Faber M,Pulmanausahakul R,Nagao K,et al.Identification of viral genomic elements responsible for rabies virus neuroin vasiveness[J].Proc Natl Acad Sci USA,2004,101(46):16328-16332.

[9] 白江,李聪.利用ELISA法对猪伪狂犬病检测的结果分析[J].中国畜牧兽医,2006,33(2):79-83.

[10] Larrous F,Gholami A,Mouhamad S,et al.Two overlapping domains of a lyssavirus matrix protein that acts on different cell death pathways[J].J Virol,2010,84(19):9897-9906.

[11] 倪娇,周绪斌,张馨玉,等.近期部分规模化猪场猪伪狂犬病野毒抗体监测情况调查[J].中国畜牧兽医,2007,34(9):47-53.

[12] Wensvoort T,Terpstra C,Pol J M,et al.Blue ear disease of pigs[J].Vet Ree,1991,1:567-574.

[13] Golding J W.Monoclonal antibodies:Principle and practice[M].New York:Academic Press,1983.

[14] 袁子国,张秀香,高胜言,等.猪伪狂犬病病毒疫苗株Bartha-K61在犬科动物体内的安全性评价及免疫实验[J].中国兽医学报,2008,10:1122-1127.

[15] 张继希,张志远,张宜娜,等.猪唾液酸黏附素氨基端结构域基因的克隆表达及其多克隆抗体的制备[J].中国畜牧兽医,2012,39(9):50-53.

[16] Finke S,Granzow H,Hurst J,et al.Intergenotypic replacement of lyssavirus matrix proteins demonstrates the role of lyssavirus M proteins in intracellular virus accumulation[J].J Virol,2010,84(4):1816-1827.

[17] Pulmanausahakul R,Li J,Schnell M J,et al.The glycoprotein and the matrix protein of rabies virus affect pathogenicity by regulating viral relication and facilitating cell-to-cell spread[J].J Virol,2008,82(5):2300-2338.

Preparation of Polyclonal Antibodies against Swine Pseudorabies Virus and Study on the Immunological Activity

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	JIAO Linlin, CHENG Yuting, WU Qingguo, WU Shuang, WU Zhi, ZHU Shanyuan, QIAN Yingjuan. Prokaryotic Expression and Polyclonal Antibody Preparation of Duck Tembusu Virus Capsid Protein [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(6): 2395-2402.
[2]	OU Yunwen, PAN Qin, WANG Yang, DAI Junfei, REN Shaoke, ZHANG Yang, ZHANG Jie. Truncated Expression of ORF2 Gene of Porcine Parvovirus Type 6 and Preparation of Its Polyclonal Antibody [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(6): 2487-2495.
[3]	WANG Xueping, ZHANG Xiaojun, LI Xiaoyue, WANG Guodong, ZHANG Fuliang, SONG Yuwei, ZHANG Mingliang, MA Lei. Prokaryotic Expression of Hexon Protein of Highly Pathogenic Fowl Adenovirus Serotype 4 and Preparation of Polyclonal Antibodies [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(5): 2053-2060.
[4]	HU Leyu, HU Xinyao, LI Ying, ZHAO Shengmiao, SHEN Fengzhen, WANG Changfa, LI Liangliang, WANG Tongtong, REN Huiying. Bioinformatics Analysis of HO-1 Gene and Preparation of Polyclonal Antibody in Dezhou Donkey [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1499-1510.
[5]	WANG Wenjie, RU Penghui, YU Chuan, DU Fuxi, CHEN Songbiao, SHANG Ke, ZHANG Chunjie, CHENG Xiangchao. Prokaryotic Expression of VP2 Protein of Luoyang Feline Panleukopenia Virus Isolate and Polyclonal Antibody Preparation [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1511-1521.
[6]	ZHANG Jingxu, XU Yu, LIANG Haiying, ZENG Zhiyong, TANG Deyuan, WANG Bin, XU Songping, WAN Juan, ZHU Yang. Phylogenetic Evolution and Epitope Analysis of Pseudorabies Virus Guizhou Isolates and Vaccine Strains [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(3): 1093-1106.
[7]	NIU Xiaojie, XU Mingguo, XIAO Li, LIU Wenhao, CHEN Chuangfu, WANG Yong. Expression and Purification of IPAJ Protein of Salmonella Pullorum, Preparation of Polyclonal Antibody and Establishment of ELISA Method [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(3): 1218-1228.
[8]	CHEN Yanxi, WANG Chen, LUO Yuan, ZHOU Yuancheng, XU Zhiwen, PENG Yuanyi, SONG Zhenhui, LIU Xiao. circRNA Expression Profile Analysis of PK-15 Cells Response to Seneca Virus A Infection [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(1): 280-289.
[9]	ZHANG Mengyang, HE Jian, LIU Yangkun, YAO Lunguang. Bioinformatics Analysis and Polyclonal Antibody Preparation of African Swine Fever Virus p37 Protein [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(1): 300-309.
[10]	ZHU Lilin, WANG Houkun, CHEN Xueyang, LIU Jing, LIANG Xiongyan, FANG Chun, YANG Yuying. Effect of Chicken IFIT5 on Replication of J Subgroup Avian Leukemia Virus Within DF-1 Cells [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(1): 310-316.
[11]	XIAO Yangyang, MA Zhongchen, LI Ruirui, CHEN Chuangfu, ZHENG Wei, WANG Yong, WANG Pengyan. Bioinformatic Analysis and Polyclonal Antibody Preparation of Brucella Secreted Protein BspI [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(8): 3112-3121.
[12]	LIU Ming, ZHANG Yongning. Prokaryotic Expression of Porcine Deltacoronavirus Nucleocapsid Protein and Preparation of Its Polyclonal Antibody [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(7): 2698-2707.
[13]	ZHOU Wei, LIU Xuping, ZHANG Yanmin, PENG Wenjuan, ZHAO Liang, TAN Wensong. Serum-free Suspension Culture of PK-15 Cells and Its Effect on the Proliferation of Porcine Circovirus Type 2 [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(6): 2397-2405.
[14]	XIE Sihao, GOU Hongchao, BIAN Zhibiao, LI Bin, CAI Rujian, ZANG Yingan, LI Chunling. Effects of MLKL Gene Knockout on Replication of Pseudorabies Virus [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(5): 1934-1941.
[15]	XU Chunmei, SHAO Mingzhu, WANG Xinyue, LIN Jiadi, GUO Jianxiong, ZHANG Xiangyin, TONG Dewen, LIANG Ruiying, ZHAO Xiaomin. Bioinformatics Analysis of African Swine Fever Virus D250R Protein and Preparation of Polyclonal Antibodies [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(12): 4745-4755.