The University of California at San Diego (UCSD), under a Federal Railroad Administration (FRA) Office of Research and Development (R&D) grant, is developing a system for high-speed and non-contact rail defect detection. A prototype using an ultrasonic air-coupled guided wave signal generation and air-coupled signal detection, paired with a real-time statistical analysis algorithm, has been realized. This system requires a specialized filtering approach based on electrical impedance matching due to the inherently poor signal-to-noise ratio of air-coupled ultrasonic measurements in rail steel. Various aspects of the prototype have been designed with the aid of numerical analyses. In particular, simulations of ultrasonic guided wave propagation in rails have been performed using a Local Interaction Simulation Approach (LISA) algorithm. The system's operating parameters were selected based on Receiver Operating Characteristic (ROC) curves, which provide a quantitative manner to evaluate different detection performances based on the trade-off between detection rate and false positive rate. Results from the first field test of the non-contact air-coupled defect detection prototype conducted at the Transportation Technology Center (TTC) in Pueblo, Colorado, in October 2014 are presented and discussed in this paper. The results indicate that the prototype is able to detect internal cracks with high reliability.