A middle ear infection is a prevalent inflammatory disease most common in the pediatric population, and its financial burden remains substantial. Current diagnostic methods are highly subjective, relying on visual cues gathered by an otoscope. To address this shortcoming, optical coherence tomography (OCT) has been integrated into a handheld imaging probe. This system can non-invasively and quantitatively assess middle ear effusions and identify the presence of bacterial biofilms in the middle ear cavity during ear infections. Furthermore, the complete OCT system is housed in a standard briefcase to maximize its portability as a diagnostic device. Nonetheless, interpreting OCT images of the middle ear more often requires expertise in OCT as well as middle ear infections, making it difficult for an untrained user to operate the system as an accurate stand-alone diagnostic tool in clinical settings. Here, we present a briefcase OCT system implemented with a real-time machine learning platform for middle ear infections. A random forest-based classifier can categorize images based on the presence of middle ear effusions and biofilms. This study demonstrates that our briefcase OCT system coupled with machine learning can provide user-invariant classification results of middle ear conditions, which may greatly improve the utility of this technology for the diagnosis and management of middle ear infections.