Development and validation of P113 gene specific polymerase spiral reaction assay for rapid detection of Mycoplasma ovipneumoniae in sheep and goats.

Respiratory infections caused by Mycoplasma ovipneumoniae remain a major challenge in sheep and goat production, resulting in reduced productivity and financial losses. Conventional diagnostic tools such as culture, PCR and ELISA are either expensive, labor intensive or unsuitable for on-site application in rural farming systems. At present, India lacks a rapid and affordable test for field detection of this pathogen. The objective of the present study is to develop Polymerase Spiral Reaction (PSR) assay to detect Mycoplasma ovipneumoniae in small ruminants. PSR primers specific for Mycoplasma ovipneumoniae P113 gene were designed for the assay. The developed PSR assay, optimized at 65 °C for 100 min, also showed a color change from deep pink to orange for positive reactions. The PSR assay was found to be highly specific to the P113 gene at low DNA concentration. When compared with PCR, the PSR assay showed 91.67% sensitivity and 92.31% specificity, making the PSR assay suitable alternative for detecting M. ovipneumoniae.