To combat antibiotic resistance, it is extremely important to select the right antibiotic by performing rapid diagnosis of pathogens. Traditional techniques require complicated sample preparation and time-consuming processes which are not suitable for rapid diagnosis. To address this problem, we used surface-enhanced Raman spectroscopy combined with machine learning techniques for rapid identification of methicillin-resistant and methicillin-sensitive Gram-positive Staphylococcus aureus strains and Gram-negative Legionella pneumophila (control group). A total of 10 methicillin-resistant S. aureus (MRSA), 3 methicillin-sensitive S. aureus (MSSA) and 6 L. pneumophila isolates were used. The obtained spectra indicated high reproducibility and repeatability with a high signal to noise ratio. Principal component analysis (PCA), hierarchical cluster analysis (HCA), and various supervised classification algorithms were used to discriminate both S. aureus strains and L. pneumophila. Although there were no noteworthy differences between MRSA and MSSA spectra when viewed with the naked eye, some peak intensity ratios such as 732/958, 732/1333, and 732/1450 proved that there could be a significant indicator showing the difference between them. The k-nearest neighbors (kNN) classification algorithm showed superior classification performance with 97.8% accuracy among the traditional classifiers including support vector machine (SVM), decision tree (DT), and naive Bayes (NB). Our results indicate that SERS combined with machine learning can be used for the detection of antibiotic-resistant and susceptible bacteria and this technique is a very promising tool for clinical applications.