Polymerase Chain Reaction and Sequence Analysis of P32 Gene of Lumpy Skin Disease Viruses Isolated During 2019 in Egypt.

Sayed, Mohamed; Kafafy, Mohamed; Mohamed, Nadine; El-Zeedy, Seham Abd El Rashid; Abbas, Ashraf Mohamed

doi:10.21608/ejvs.2023.191819.1462

Polymerase Chain Reaction and Sequence Analysis of P32 Gene of Lumpy Skin Disease Viruses Isolated During 2019 in Egypt.

Document Type : Original Article

Authors

¹ *Agriculture Research Centre. *Veterinary Serum and Vaccine Research Institute. *Genetic Engineering Research Department.

² *Agriculture Research Centre. *Veterinary Serum and Vaccine Research Institute. *Pox Virus Vaccine Research Department.

10.21608/ejvs.2023.191819.1462

Abstract

Lumpy skin disease (LSD) is one of the important viral diseases affecting cattle herds Not only in Egypt but all over the MENA region, Africa and Asia as well, the current Molecular study based on molecular detection and sequence analysis of Lumpy skin Disease virus using the P32 gene which is very conservative gene not only for LSDV But for the whole Capri pox group viruses including the sheep and goat poxviruses, Skin lesion from cattle showing the typical clinical picture of the disease from Sharqia governorate during summer 2019 gives positive PCR band, at the expected size. Which were about 1185 bp where the selected primers were up stream of the P32 gene open reading frame By 213 bp till the end of the coding sequence which spans bout 969 bp which is the Coding sequence of the gene of interest, the PCR product of the target product is 1185 bp Including the coding region of the P32 gene sequenced and submitted to gene bank accession number#OL423259, this primer could be used as universal primer for all Capripox group, the current study used PCR-RFLP to differentiate between the sheep poxvirus and Lumpy Skin Disease virus by Specific characteristic pattern for each virus and could be used as a tool of Differentiation between these two viruses, the used bioinformatics tools revealed Presence of two different signature residues very characteristic for either the local Isolate VSVRI/Sharqia/2019 and Lumpy Skin Disease virus in general and the Romanian sheep pox reference strain used in this study which assures the slight Difference in the antigenicity of the two viruses even though the great homology of The two viruses at the nucleotide sequence level while at the amino acid level residues vary Slightly in length in sheep pox virus is about 323 residues while in lumpy skin disease The amino acid residues of the P32 is only 322 residues due to insertion of extra aspartate residue at position 55 which is not present in Lumpy Skin Disease virus and even goat pox virus Which is related to the same Capri group which reveals slight differences between the sheep Poxvirus and lumpy skin Disease virus in antigensity so this insertion in SPV make the slight difference in length of the coding nucleotide sequence between the two viruses using HinfI restriction enzyme is very unique in the current study and could be used as a sensitive rapid tool to differentiate between the two viruses as it has two recognition sites of digestion in SPV while in LSDV has only one recognition site this may be due to the difference in the arrangement of the nucleotide along the length of the coding sequence of P32 due to the insertion of extra 3 nucleotide in SPV which is not present in LSDV.

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