| Popis: |
Lumpy skin disease (LSD) is an exotic disease to India, affecting cattle and buffalo. LSD originated in Africa, where many domesticated and wild ruminant species are affected. LSD was first recorded in India from five districts of Odisha state during August 2019. Subsequently, LSD outbreaks were reported in more than fourteen states of India. The aim of the study was to develop a sensitive SYBR green real-time polymerase chain reaction assay for the diagnosis of LSD in the tissue biopsy samples. Tissue biopsy samples collected from cattle showing signs of LSD (n=8) were subjected to conventional gel-based PCR and SYBR green real-time PCR using the primer sets specific for the gene of viral attachment protein (VP32). Seven of eight tissue biopsy samples showed the positive amplification of 192bp product size in conventional gel-based PCR with primers recommended by OIE. All the eight tissue biopsy samples were positive in SYBR green real-time PCR assay, based on the same OIE primers, and the threshold value (Ct) values varied from 19.14 to 23.01. The seven PCR amplicons were further sequenced to confirm the identification of LSDV VP32 gene. Further, the sequences were accessioned in GenBank and their phylogenetic analysis revealed close phylogenetic relation to a partial LSDV p32 gene sequence from Egypt. The developed SYBR green real-time PCR assay is a cost-effective, specific, sensitive and rapid diagnostic assay for detection of LSD and quantification of LSD viral load in the tissue biopsy samples. |
