Mastitis is one of the most common infectious diseases that causes the financial losses for dairy farms. The primary step between determining the etiology of the disease and finding a treatment is accurate diagnosis. Researchers make an effort to develop sensitive, specific, rapid, and cost-effective diagnostic methods for bovine subclinical mastitis caused by staphylococcus aureus to solve problems with current diagnostic methods. There are currently new developments in the use of nanotechnology-based techniques for colorimetric, immunological, and molecular detection to design biosensors for the accurate, selective, and functional detection of biomolecules. Nanoparticles are good for replacement and incorporation with other typical traditional subclinical mastitis diagnostic methods due to their nanoparticle size and adaptable physicochemical features including; electrical, electrochemical, optical, and magnetic properties. Hence, the present review discusses the applications and the role of nanotechnology in the diagnosis of subclinical staphylococcal aureus bovine mastitis.