The present chapter deals with conventional methods of detection and identification of microorganisms with a prime focus on food microbiology. It provides a brief overview of the different detection methods of microorganisms, such as standard plate count method, most probable number count and direct microscopic count. Similarly, the identification methods as mentioned in this chapter are a series of biochemical tests. These suggest the presence of certain enzyme systems in the microorganism or metabolic by-product in the culture medium and using this information, the target microorganism can be identified. The biochemical tests (i.e. IMViC test, catalase test, oxidase test, urease test and nitrate and nitrite reduction test), discussed in this chapter are widely used by the microbiological laboratory. Apart from the tests mentioned above, there are, however, different kinds of culture-dependent biochemical tests developed over the years to detect the presence of some specific enzyme system or metabolic by-products.