Background: Lumpy skin disease (LSD) presents a significant threat to the global livestock industry, with potential implications for both animal health and economic stability. The recombinant A27L protein, derived from the lumpy skin disease Virus (LSDV), is a candidate for both diagnostic applications and vaccine development. 
 
Methods: In this study, we successfully cloned, expressed and purified the A27L protein using an E. coli expression system. A 447 bp fragment of the A27L gene was amplified from LSDV DNA, cloned into the pJET1.2/blunt vector and subcloned into the pQE30 expression vector. Expression of the recombinant protein was induced in E. coli M15 cells, yielding a 17.7 kDa protein, which was subsequently purified using nickel affinity chromatography. Additionally, a real-time PCR assay was performed for the differentiation of LSDV from other related poxviruses, enhancing the specificity of diagnostic testing. 
 
Result: Preliminary diagnostic validation of A27L protein was conducted using a DOT ELISA with LSDV-positive field serum sample, showing promising results. While the initial findings suggest the potential of the recombinant A27L protein for diagnostic use, further validation, including larger sample sizes and comparisons with existing diagnostic methods, is necessary to fully establish its effectiveness as a diagnostic tool and vaccine candidate. In addition, real-time PCR assay demonstrated specific detection and differentiation of LSDV from other related poxviruses. 
&nbsp;

Recombinant A27L Protein and Real-time PCR-based Detection of Lumpy Skin Disease Virus: A Step Toward Enhanced Diagnosis and Vaccine Research