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Differentiation of Sheeppox and Goatpox Viruses by Polymerase Chain Reaction-Restriction Fragment Length Polymorphism

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Virologica Sinica 2012年第6期27卷 353-359页

作者： Gnanavel Venkatesan Vinayagamurthy Balamurugan revaniah yogisharadhya Amit Kumar Veerakyathappa Bhanuprakash Division of Virology Indian Veterinary Research InstituteNainital(Distt.)Mukteswar 263138UttarakhandIndia Project Directorate on Animal Disease Monitoring and Surveillance H A FarmHebbalBangalore 560024KarnatakaIndia Indian Veterinary Research Institute H A FarmHebbalBangalore 560024KarnatakaIndia

In the present study,the partial gene sequences of P32 protein,an immunogenic envelope protein of Capripoxviruses (CaPV),were analyzed to assess the genetic relationship among sheeppox and goatpox virus isolates,and restriction enzyme specific PCR-RFLP was developed to differentiate CaPV strains.A total of six goatpox virus (GTPV) and nine sheeppox virus (SPPV) isolates of Indian origin were included in the sequence analysis of the attachment *** sequence analysis revealed a high degree of sequence identity among all the Indian SPPV and GTPV isolates at both nucleotide and amino acid *** analysis showed three distinct clusters of SPPV,GTPV and Lumpy skin disease virus (LSDV) ***,multiple sequence alignment revealed a unique change at G120A in all GTPV isolates resulting in the formation of Dra I restriction site in lieu of EcoR I,which is present in SPPV isolates *** change was unique and exploited to develop restriction enzyme specific PCR-RFLP for detection and differentiation of SPPV and GTPV *** optimized PCR-RFLP was validated using a total of fourteen (n=14) cell culture isolates and twenty two (n=22) known clinical samples of *** Restriction Enzyme specific PCR-RFLP to differentiate both species will allow a rapid differential diagnosis during CaPV outbreaks particularly in mixed flocks of sheep and goats and could be an adjunct/supportive tool for complete gene or virus genome sequencing methods.

关键词：山羊痘病毒限制性片段长度多态性聚合酶链反应 PCR-RFLP技术分化限制性内切酶基因序列系统发育分析

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