Abstract: The microbiology assay is the primary detection method of national standards as traditional classical method. Although the microbiological method has the characteristics of long time consuming,low accuracy and poor repeatability,it still needs to be determined by this method for vitamins with low content and lower detection limit. This method is mainly used in the detection of vitamins in infant foods,but few studies for other food categories. This paper described the applications and research progress of microbiological assay for vitamin B₁₂ and folic acid in foods. The main contents contain controlling and optimizing the key points,of national standard method and establishing the high throughput and fast detection method,which will provide some references for the two vitamins determination with microbiological methods. The microbiological methods of VB₁₂ and folic acid in food are at the stage of optimization and development,which can promote the standardization and utilization of methods. With the development of new technology,it will be the future trend of vitamins detection in high throughput and multicomponent simultaneous detection.