Abeer M Abdalhamed¹*, Eman S Ibrahim², Dina Y H EL-Shafey³ and Gamil S G Zeedan¹

Mycoplasma bovis (M. bovis) infection is causing substantial economic losses to the global dairy industry including Egypt through its role as a major mastitis pathogen in cattle. The present study was aimed to evaluate loop-mediated isothermal amplification methods (LAMP) and real-time quantitative PCR (rt-qPCR) assays for detecting M. bovis in milk samples from cows (n=30) and buffaloes (n=20) with mastitis across different governorates in Egypt between January and March 2023. Both assays were compared to traditional culture and counter-immunoelectrophoresis (CIEP) methods. The results of the culture methods were 16 (32%), CIEP was 10 (20%), PCR was 16 (32%), LAMP was 18 (36%), and rt-qPCR based on Syber Green was 19 (38%), respectively. The sensitivity of rt-qPCR has the highest sensitivity (100%), specificity (91.17%), and accuracy (100%) (kappa coefficient of 0.856). While LAMP has sensitivity 93.75%, specificity 91.17%, and accuracy 98% (kappa coefficient of 0.811). However, the lowest detection rate was found in culture methods and CIEP. Our research effectively utilized LAMP and Sybr Green-based rt-qPCR assays for the rapid detection of M. bovis in bovine milk samples. However, further validation is necessary with a larger sample size.