Year: 2025 | Month: February | Volume 15 | Issue 1
Inactivation of Lumpy Skin Disease Virus Isolate Obtained from the Field Outbreaks using Binary Ethylenimine
Bommena Sushma
M. Srinivas
L. Gopala
Kalyani Putty and G.B. Manjunatha Reddy
DOI:10.30954/2277-940X.01.2025.7
Abstract:
Present study was aimed to isolate and inactivate the lumpy skin disease virus from the field outbreaks, clinically suspected
samples like skin lesions, skin scabs, skin nodules and blood were collected from the affected bovine species. The samples were
homogenised and processed, DNA was isolated and they were initially amplified for Capripoxvirus specific partial P32 gene,
followed by LSDV-specific EEV glycoprotein gene-LSDV126 using respective forward and reverse primers. The PCR positive
samples are subjected for isolation and propagation MDBK cells. Once the CPE was noticed in infected flasks, the cells were
lysed by three cycles of freeze and thaw, the culture fluid was collected and stored at -20°C. The infected cell culture fluid was
subjected for molecular detection by conventional PCR by primers targeting LSDV126 gene which is specific for LSDV. After
molecular confirmation the viral isolates were assayed for titres and the infectivity titre was calculated by Reed and Muench
(1938) and subjected for inactivation using BEI.
Highlights
- Detection of LSD virus in skin lesions, nodules, scabs and blood using Real time PCR.
- Isolation of LSD virus using MDBK cell lines.
- Inactivation of LSD virus using Binary Ethylenimine (BEI).
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