Fish is an essential source of many nutrients necessary for human health. However, the deliberate mislabeling of similar fish fillet types is common in markets to make use of the relatively high price difference. This is a type of explicit food adulteration. In the present work, spectrochemical analysis and chemometric methods are adopted to disclose this type of fish species cheating. Laser-induced breakdown spectroscopy (LIBS) was utilized to differentiate between the fillets of the low-priced tilapia and the expensive Nile perch. Furthermore, the acquired spectroscopic data were analyzed statistically using principal component analysis (PCA) and artificial neural network (ANN) showing good discrimination in the PCA score plot and a 99% classification accuracy rate of the implemented ANN model. The recorded spectra of the two fish indicated that tilapia has a higher fat content than Nile perch, as evidenced by higher CN and C2 bands and an atomic line at 247.8 nm in its spectrum. The obtained results demonstrated the potential of using LIBS as a simple, fast, and cost-effective analytical technique, combined with statistical analysis for the decisive discrimination between fish fillet species.