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

• Published in: Virologica Sinica Vol. 27; no. 6; pp. 352 - 358
• Main Authors: Venkatesan, Gnanavel, Balamurugan, Vinayagamurthy, Yogisharadhya, Revaniah, Kumar, Amit, Bhanuprakash, Veerakyathappa
• Format: Journal Article
• Language: English
• Published: Heidelberg Springer-Verlag 01-12-2012; SP Wuhan Institute of Virology, CAS; KeAi Publishing Communications Ltd; Division of Virology, Indian Veterinary Research Institute, Nainital(Distt.), Mukteswar 263138, Uttarakhand, India%Division of Virology, Indian Veterinary Research Institute, Nainital(Distt.), Mukteswar 263138, Uttarakhand, India; Project Directorate on Animal Disease Monitoring and Surveillance, H A Farm, Hebbal, Bangalore 560024,Karnataka, India%Division of Virology, Indian Veterinary Research Institute, Nainital(Distt.), Mukteswar 263138, Uttarakhand, India; Indian Veterinary Research Institute, H A Farm, Hebbal, Bangalore 560024, Karnataka, India
• Subjects: Amino acid sequence; Amino acids; Animals; Biochemistry; Biomedical and Life Sciences; Biomedicine; Capripoxvirus; Capripoxvirus - classification; Capripoxvirus - genetics; Capripoxvirus - isolation & purification; Cell culture; Cluster Analysis; Differential diagnosis; Differentiation; DNA, Viral - chemistry; DNA, Viral - genetics; Env protein; Envelope protein; Enzymes; Gene polymorphism; genes; Genetic relationship; genetic relationships; Genomes; Goatpox virus; Goats; Immunogenicity; India; Lumpy skin disease; Lumpy skin disease virus; Medical Microbiology; Microbial Genetics and Genomics; Microbiology; Molecular Sequence Data; Nucleotide sequence; nucleotide sequences; Oncology; p32 Protein; PCR-RFLP技术; Phylogeny; Point Mutation; Polymerase chain reaction; Polymerase Chain Reaction - methods; Polymorphism, Genetic; Polymorphism, Restriction Fragment Length; Research Article; Restriction fragment length polymorphism; sequence alignment; Sequence analysis; Sequence Analysis, DNA; Sheep; Sheeppox virus; Skin diseases; Strains (organisms); Viral envelope proteins; Viral Proteins - genetics; Virology; Viruses; 分化; 基因序列; 山羊痘病毒; 系统发育分析; 聚合酶链反应; 限制性内切酶; 限制性片段长度多态性; India; Sheeppox; Differential diagnosis; PCR-RFLP; Goatpox
• ISSN: 1674-0769; 1995-820X
• DOI: 10.1007/s12250-012-3277-2

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 gene. The sequence analysis revealed a high degree of sequence identity among all the Indian SPPV and GTPV isolates at both nucleotide and amino acid levels. Phylogenetic analysis showed three distinct clusters of SPPV, GTPV and Lumpy skin disease virus （LSDV） isolates. Further, 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 studied. This change was unique and exploited to develop restriction enzyme specific PCR-RFLP for detection and differentiation of SPPV and GTPV strains. The optimized PCR-RFLP was validated using a total of fourteen （n=14） cell culture isolates and twenty two （n=22） known clinical samples of CaPV. The 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.