Attenuation of Virulent PorcineReproductive andRespiratorySyndrome Virus Strain CH-1a and Genetic Variation of ORF5 Gene

doi:10.1016/S1671-2927(00)8741

Journal of Integrative Agriculture ›› 2012, Vol. 12 ›› Issue (12): 2035-2042.DOI: 10.1016/S1671-2927(00)8741

Attenuation of Virulent PorcineReproductive andRespiratorySyndrome Virus Strain CH-1a and Genetic Variation of ORF5 Gene

CAI Xue-hui, WUGuo-jun, LIU Yong-gang, LIU Guang-qing, SHI Wen-da, WANG Shu-jie, MA Ping, LI Cheng-jun , HAN Wen-yu

1.College of Animal Science and Veterinary Medicine, Jilin University, Changchun 130062, P.R.China
2.State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences,
Harbin 150001, P.R.China
3.Division of Avian Infectious Diseases, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai
200241, P.R.China

收稿日期:2012-06-11 出版日期:2012-12-01 发布日期:2012-12-18
通讯作者: Correspondence HAN Wen-yu, Tel: +86-431-87836001, Fax: +86-431-87836029, E-mail: hanwy@jlu.edu.cn
作者简介:CAI Xue-hui, Tel: +86-451-51997183, E-mail: aci139@sina.com
基金资助:
This work was supported by the National High Technology Research and Development Program of China (2011AA10A213), the Key Technology R&D Program of Harbin, China (2010AA6AN083), the Excellent Youth Foundation of Heilongjiang Province of China (JC201020).

Attenuation of Virulent PorcineReproductive andRespiratorySyndrome Virus Strain CH-1a and Genetic Variation of ORF5 Gene

CAI Xue-hui, WUGuo-jun, LIU Yong-gang, LIU Guang-qing, SHI Wen-da, WANG Shu-jie, MA Ping, LI Cheng-jun , HAN Wen-yu

1.College of Animal Science and Veterinary Medicine, Jilin University, Changchun 130062, P.R.China
2.State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences,
Harbin 150001, P.R.China
3.Division of Avian Infectious Diseases, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai
200241, P.R.China

Received:2012-06-11 Online:2012-12-01 Published:2012-12-18
Contact: Correspondence HAN Wen-yu, Tel: +86-431-87836001, Fax: +86-431-87836029, E-mail: hanwy@jlu.edu.cn
About author:CAI Xue-hui, Tel: +86-451-51997183, E-mail: aci139@sina.com
Supported by:
This work was supported by the National High Technology Research and Development Program of China (2011AA10A213), the Key Technology R&D Program of Harbin, China (2010AA6AN083), the Excellent Youth Foundation of Heilongjiang Province of China (JC201020).

摘要/Abstract

摘要： To develop a modified live vaccine (MLV) against porcine reproductive and respiratory syndrome virus (PRRSV), virulent CH-1a strain was attenuated by serial passages up to 130 passage (P130) in Marc-145 cells. The virulence and immune efficacy of the attenuated CH-1a were evaluated in pigs. The results showed that animals inoculated with P130 did not develop any clinical sign of the disease, but produced rapid and effective humoral immune responses against PRRSV challenge, indicating that attenuated CH-1a P130 is the candidate as the effective vaccine against PRRSV. To define the potential mutations in the attenuated CH-1a genome, we sequenced and analyzed the ORF5 gene of CH-1a strain of different passages (P39, P55, P65, P70, P85, P100, P115, P120, P125, and P130) and found that three mutations (C5Y, H38Q and L146Q) which may be related with the attenuation of CH-1a. In addition, we also found a unique restriction enzyme site (TspEI) in the ORF5 gene of attenuated CH-1a, which can be used as a genetic marker to distinguish original and attenuated CH-1a.

关键词: attenuation, porcine reproductive and respiratory syndrome virus, genetic variation, ORF5 gene

Abstract: To develop a modified live vaccine (MLV) against porcine reproductive and respiratory syndrome virus (PRRSV), virulent CH-1a strain was attenuated by serial passages up to 130 passage (P130) in Marc-145 cells. The virulence and immune efficacy of the attenuated CH-1a were evaluated in pigs. The results showed that animals inoculated with P130 did not develop any clinical sign of the disease, but produced rapid and effective humoral immune responses against PRRSV challenge, indicating that attenuated CH-1a P130 is the candidate as the effective vaccine against PRRSV. To define the potential mutations in the attenuated CH-1a genome, we sequenced and analyzed the ORF5 gene of CH-1a strain of different passages (P39, P55, P65, P70, P85, P100, P115, P120, P125, and P130) and found that three mutations (C5Y, H38Q and L146Q) which may be related with the attenuation of CH-1a. In addition, we also found a unique restriction enzyme site (TspEI) in the ORF5 gene of attenuated CH-1a, which can be used as a genetic marker to distinguish original and attenuated CH-1a.

CAI Xue-hui, WUGuo-jun, LIU Yong-gang, LIU Guang-qing, SHI Wen-da, WANG Shu-jie, MA Ping, LI Cheng-jun , HAN Wen-yu. Attenuation of Virulent PorcineReproductive andRespiratorySyndrome Virus Strain CH-1a and Genetic Variation of ORF5 Gene[J]. Journal of Integrative Agriculture, 2012, 12(12): 2035-2042.

CAI Xue-hui, WUGuo-jun , LIU Yong-gang, LIU Guang-qing, SHI Wen-da, WANG Shu-jie, MA Ping, LI Cheng-jun , HAN Wen-yu. Attenuation of Virulent PorcineReproductive andRespiratorySyndrome Virus Strain CH-1a and Genetic Variation of ORF5 Gene[J]. Journal of Integrative Agriculture, 2012, 12(12): 2035-2042.

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Attenuation of Virulent PorcineReproductive andRespiratorySyndrome Virus Strain CH-1a and Genetic Variation of ORF5 Gene

Attenuation of Virulent PorcineReproductive andRespiratorySyndrome Virus Strain CH-1a and Genetic Variation of ORF5 Gene

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