猪圆环病毒2型不同基因型毒株感染性克隆的构建及拯救毒株的体外生物学特性

doi:10.3864/j.issn.0578-1752.2011.23.019

Abstract

Abstract: 【Objective】 Different genotypes (PCV2a, PCV2b and PCV2d) of porcine circovirus type 2 (PCV2) are present in clinical PCV2 infections in China, and it is necessary to elucidate the pathogenic difference among different genotypes of PCV2. 【Method】Representative strains of different genotypes of PCV2 were constructed by infectious molecular clone and biologic characterization of the rescued viruses were identified in vitro. 【Result】 Four infectious clones for different genotypes of PCV2 were constructed and four viruses of different genotypes were rescued after transfection into PK15 cells. The rescued viruses were PCV2 after identified by nucleotide sequence analysis, morphology of the viruses and immunoperoxidase monolayer assay (IPMA). The rescued viruses propagated stably after consecutive incubation for 10 passages, and virus propagation reached up to peak at 72 h post infection (PI), and the virus titers were up to 105.0TCID50/mL. By using neutralizing 1D2 monoclonal antibody (mAb) of PCV2, the antigen capture ELISA showed that only rescued PCV2a/rCL had reactivity with 1D2 mAb, and however, another three rescued strains (PCV2b/rYJ, PCV2b/rJF and PCV2d/rBDH) did not, which indicated the antigenic difference among the rescued viruses of different genotypes.【Conclusion】Conclusion could be draw from this study that PCV2 prevailing has differences in genomic and ORF2 gene length and antigen in swine herds. Four representative clones for different genotypes were constructed and rescued, which will facilitate further studies on the pathogenic difference resulted from different genotypes of PCV2.

Key words: porcine circovirus type 2 (PCV2), different genotypes, infectious molecular clone, rescue virus, biologic characteristic

GUO Long-Jun, LU Yue-Hua, HUANG Li-Ping, WEI Yan-Wu, LIU Chang-Ming. Construction of Infectious Clone for Different Genotype Strains of Porcine Circovirus Type 2 and Characterization of the Rescued Viruses in vitro[J].Scientia Agricultura Sinica, 2011, 44(23): 4918-4925.

0
/ / Recommend

Add to citation manager EndNote|Reference Manager|ProCite|BibTeX|RefWorks

URL: https://www.chinaagrisci.com/EN/10.3864/j.issn.0578-1752.2011.23.019

https://www.chinaagrisci.com/EN/Y2011/V44/I23/4918

References

[1]Mankertz A, Domingo M, Folch J M, Le Cann P, Jestin, A, Segales, J, Chmielewicz B, Plana-Duran J, Soike D. Characterization of PCV2 isolates from Spain, Germany and France. Virus Research, 2000, 66 (1): 65-77.

[2]Ellis J, Clark E, Haines D, West K, Krakowka S, Kennedy S, Allan G M. Porcine circovirus-2 and concurrent infections in the field. Veterinary Microbiology, 2004, 98: 159-163.

[3]Wellenberg G J, Stockhofe Z N, Boersma W J, De Jong M F, Elbers A R W. The presence of co-infections in pigs with clinical signs of PMWS in the Netherlands: case-control study. Research Veterinary Science, 2004, 77 (2): 177-184.

[4]Liu C M, Wei Y W, Zhang C F, Lu Y H, Kong X G. Construction and characterization of porcine circovirus type 2 carrying a genetic marker strain. Virus Research, 2007, 127: 95-99.

[5]Mankertz A, Hillenbrand B. Replication of porcine circovirus type 1 requires two proteins encoded by the viral rep gene. Virology, 2001, 279: 429-438.

[6]Cheung A K. Transcriptional analysis of porcine circovirus type 2. Virology, 2003, 305: 168-180.

[7]Cheung A K. Comparative analysis of the transcriptional patterns of pathogenic and nonpathogenic porcine circoviruses. Virology, 2003, 310: 41-49.

[8]Nawagitgul P, Morozov I, Bolin S R, Harms P A, Sorden S D, Paul P S. Open reading frame 2 of porcine circovirus type 2 encodes a major capsid protein. Journal of General Virology, 2000, 81: 2281-2287.

[9]Liu J, Chen I, Kwang J. Characterization of a previously unidentified viral protein in porcine circovirus type 2-infected cells and its role in virus-induced apoptosis. Journal of Virology, 2005, 79: 8262-8274.

[10]Liu J, Chen I, Du Q, Chua H, Kwang J. The ORF3 protein of porcine circovirus type 2 is involved in viral pathogenesis in vivo. Journal of Virology, 2006, 80: 5065-5073.

[11]Wen L B, Guo X, Yang H C. Genotyping of porcine circovirus type 2 from a variety of clinical conditions in china. Veterinary Microbiology, 2005, 110: 141-146.

[12]刘新文, 曹胜波, 郭东春, 姚清侠, 陈焕春. 猪Ⅱ型圆环病毒(PCV2)Henan株的分离与全基因组序列分析. 中国兽医学报, 2006, 26(4): 357-359.

Liu X W, Cao S B, Guo D C, Yao Q X, Chen H C. Isolation and genomic sequence analysis of porcine circovirus type 2(PCV2) Henan strain. Chinese Journal of Veterinary Medicine, 2006, 26(4): 357-359. (in Chinese)

[13]Fenaux M, Halbur P G, Gill M, Toth T, Meng X J. Genetic characterization of type 2 porcine circovirus(PCV2) from pigs with postweaning multisystemic wasting syndrome in different geographic regions of Northern America and development of a differential PCR-restriction fragment length polymorphism assay to detect and differentiate between infectious with PCV-1 and PCV-2. Journal of Clinical Microbiology, 2000, 38 (7): 2494-2503.

[14]Choi K S, Chae J S. Genetic characterization of porcine circovirus type 2 in Republic of Korea. Research in Veterinary Science, 2008, 84: 497-501.

[15]Grau R L, Crisci E, Sibila M, Lopez S S, Nofrarias M, Cortey M, Fraile L, Olvera A, Segalés J. A proposal on porcine circovirus type 2 (PCV2) genotype definition and their relation with postweaning multisystemic wasting syndrome (PMWS) occurrence. Veterinary Microbiology, 2008, 128: 23-35.

[16]Wang F, Guo X, Ge X N, Wang Z T, Chen Y H, Cha Z L, Yang H C. Genetic variation analysis of Chinese strains of porcine circovirus type 2. Virus Research, 2009, 145: 151-156.

[17]Ciacci J R, Simon N L, Pinto L S, Viancelli A, Fernandes L T, Hayashi M, Dellagostin O A, Esteves P A. Detection of porcine Circovirus type 2 (PCV2) variants PCV2-1 and PCV2-2 in Brazilian pig population. Research in Veterinary Science, 2009, 87: 157-160.

[18]Dupont K, Nielsen E O, Bakbo P, Larsen L E. Genetic analysis of PCV2 isolates from Danish archives and a current PMWS case-control study supports a shift in genotypes with time. Veterinary Microbiology, 2008, 128: 56-64.

[19]Grau R L, Fraile L, Segalés J. Recent advances in the epidemiology, diagnosis and control of diseases caused by porcine circovirus type 2. The Veterinary Journal, 2010 (in press).

[20]刘长明, 张超范, 危艳武, 谭斌, 陆月华, 谷守林. 猪圆环病毒2型细胞培养适应毒株的培育和鉴定. 中国预防兽医学报, 2006, 28(3): 248-252.

Liu C M, Zhang C F, Wei Y W, Tan B, Lu Y H Gu S L. Cultivation and identification of porcine circovirus type 2(PCV2) cell culture-adapted strain. Chinese Journal of Preventive Veterinary Medicine, 2006, 28(3): 248-252. (in Chinese)

[21]郭龙军, 陆月华, 危艳武, 黄立平, 刘长明. 我国部分地区猪圆环病毒2型遗传变异分析. 中国预防兽医学报, 2009, 31(11): 856-859.

Guo L J, Lu Y H, Wei Y W, Huang L P, Liu C M. Genetic variation analysis of porcine circovirus type 2 isolates from different regions of China. Chinese Journal of Preventive Veterinary Medicine, 2009, 31(11): 856-859. (in Chinese)

[22]黄立平, 刘长明, 危艳武, 张朝霞, 陆月华, 郭龙军. 抗猪圆环病毒2型Cap蛋白中和活性单克隆抗体的制备及鉴定. 中国预防兽医学报, 2009, 31(2): 132-136.

Huang L P, Liu C M, Wei Y W, Zhang Z X, Lu Y H, Guo L J. Preparation and identification of neutralizing monoclonal antibody to the recombinant capsid protein of porcine circovirus type 2. Chinese Journal of Preventive Veterinary Medicine, 2009, 31(2): 132-136. (in Chinese)

[23]刘长明, 张超范, 危艳武, 张朝霞, 袁婧. 猪圆环病毒2型免疫过氧化物酶单层细胞试验抗体检测试剂盒的研制及应用. 中国预防兽医学报, 2007, 29(8): 621-624.

Liu C M, Zhang C F, Wei Y W, Zhang Z X, Yuan J. Development and application of an immunoperoxidase monolayer assay based kit for detection of porcine circovirus type 2. Chinese Journal of Preventive Veterinary Medicine, 2007, 29(8): 621-624. (in Chinese)

[24]Olvera A, Cortey M, Segalés J. Molecular evolution of porcine circovirus type 2 genomes:Phylogeny and clonality. Virology, 2007, 357: 175-185.

[25]Knell S, Willems H, Hertrampf B, Reiner G. Comparative genetic characterization of porcine circovirus type 2 samples from German wild boar populations. Veterinary Microbiology, 2005, 109: 169-177.

[26]郗鑫, 陈焕春, 陈华平, 曹胜波, 琚春梅. 猪Ⅱ型圆环病毒全基因组的克隆及感染性鉴定. 微生物学报, 2004, 44(2): 172-176.

Xi X, Chen H C, Chen H P, Cao S B, Ju C M. Cloning of full length genome of porcine circovirus type 2 and identification of the infectivity. Acta Microbiologica Sinica, 2004, 44(2): 172-176. (in Chinese)

Metrics

Viewed

Full text

Abstract

Cited

Shared

Discussed

Comments

Recommended 0

No Suggested Reading articles found!

Construction of Infectious Clone for Different Genotype Strains of Porcine Circovirus Type 2 and Characterization of the Rescued Viruses in vitro

PDF

Abstract

Cite this article

share this article

References

Related Articles 4

Metrics

Comments

Recommended 0

[1]	DAI Hai-Fang, WU Hui, A Man-Gu-Li-?Mai-Mai-Ti-A-Li, WANG Li-Hong, MAI Mai-Ti-?A-Pi-Zi, ZHANG Ju-Song. Analysis of Salt-Tolerance and Determination of Salt-Tolerant Evaluation Indicators in Cotton Seedlings of Different Genotypes [J]. Scientia Agricultura Sinica, 2014, 47(7): 1290-1300.
[2]	SUN Yun,HUA Li-zhong,CHEN Geng,ZHANG Shu-xia . Changes of Acute Phase Proteins Profile in PCV2 Experimentally Infected Piglets [J]. Scientia Agricultura Sinica, 2010, 43(19): 4116-4122 .
[3]	. Rescue of Infectious Foot-and-Mouth Disease Virus in vivo from Full-Length cDNA Clone of Asia1 Type [J]. Scientia Agricultura Sinica, 2009, 42(2): 688-693 .
[4]	,,,. Study on the biologic characteristic of rolC-transgenic Malus robusta Rehd in vitro [J]. Scientia Agricultura Sinica, 2006, 39(12): 2563-2569 .