Both dead and living bacteria can affect food quality. In recent years, the development of microbiological testing has undergone a series of progressions from traditional methods—including microbial cultivation, physiological and biochemical testing, instrument analysis, and immunology—to molecular biological detection. Reviewing its developments, it can be seen that the detection is becoming more rapid, sensitive, precise, integrated, and low cost. Of course, when all methods of microbial detection are compared, only living bacteria that are detected by traditional methods could yield false-negative results, especially if there are dormant bacteria. But with molecular methods, all microorganisms can be detected. With the development of technology, a lot of focus has been given to identification and detection technology for dead and living bacteria, which has massive potential and important significance in the area of microorganisms. There exist prospects of novel detection techniques for discrimination of viable and nonviable microorganisms, especially some more sensitive, precise, and comprehensive ones.