Polymerase chain reaction based lumpy skin disease diagnostic test using viral p32 gene

Abstract

Infectious diseases of animals pose a significant threat to animal health and food security. Recently, several viruses have created significant losses in animal life, economy of India and across the globe. Lumpy Skin Disease (LSD), caused by Lumpy skin disease virus (LSDV) has become rampant among cattle in several northern states of India with dire consequences. LSDV is grouped under genus Capripoxvirus of family Poxviridae. Capripoxviruses are cross-reactive within the genus, therefore sheep pox or goat pox vaccines have been used to provide cross protection against lumpy skin disease virus. The diagnosis of LSDV relies on ELISA, FAT, western blotting, Polymerase chain reaction, etc. PCR being the most preferred diagnostic test for LSDV, however the method prescribed by OIE lack specificity. Therefore, the investigation was initiated with the objective to enhance sensitivity and specificity of the PCR method for diagnosis. It included optimization of PCR based method which is able to detect LSD and its differentiation with Goat Pox and Sheep pox Viruses. The method was validated by isolation of DNA from 18 samples of LSDV affected animals, 2 samples from apparently healthy animals, goat pox, sheep pox and camel pox virus. The method efficiently detected the organism from all the 18 sample of LSDV affected animals and reacted negatively with 2 samples from healthy animals, sheep pox, goat pox and camel pox. The method offered remarkable limit of detection of 17.78 pg of DNA template.