Throughout history, diseases of plants, animals, and humans transmitted by microscopic airborne particles have had a significant impact on society and the economy. Since the nineteenth century, these particles have been collected using specialized air-sampling equipment, but it has frequently been unable to identify them with accuracy. The real-time PCR technique combines the efficiency of traditional PCR with the generation of a particular fluorescent signal. It measures the kinetics of the reaction during the initial stages of PCR and allows for the quantification of specific targets in a variety of environmental samples. Since PCR products fall into the categories of amplicon sequence non-specific and sequence-specific procedures, there is an expanding variety of chemistries available for their detection. This chapter describes the exciting new possibilities for identifying and counting airborne plant pathogens, including bacteria, fungi, and viruses, by combining real-time PCR with air monitoring. As a vital tool in research laboratories, this technique has emerged as the gold standard for the early detection of airborne pathogens. Furthermore, using these techniques, one can forecast the likelihood of unforeseen disease outbreaks in the air by detecting genetic alterations in pathogen populations that make management impossible or increases in pathogen inoculum. Policymakers, medical experts, and the agriculture sector can all benefit from these forecasts by using them as a guide when developing plans to reduce the likelihood of severe pandemics.