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This work presents the medium-resolution ($R \sim 1,500$) spectroscopic follow-up of 522 low-metallicity star candidates from the Southern Photometric Local Universe Survey (S-PLUS). The objects were selected from narrow-band photometry, taking advantage of the metallicity-sensitive S-PLUS colors. The follow-up observations were conducted with the Blanco and Gemini South telescopes, using the COSMOS and GMOS spectrographs, respectively. The stellar atmospheric parameters (T$_{\rm eff}$, log$g$, and [Fe/H]), as well as carbon and $α$-element abundances, were calculated for the program stars in order to assess the efficacy of the color selection. Results show that $92^{+2}_{-3}\%$ of the observed stars have [Fe/H]$\leq -1.0$, $83^{+3}_{-3}\%$ have [Fe/H]$\leq -2.0$, and $15^{+3}_{-3}\%$ have [Fe/H]$\leq -3.0$, including two ultra metal-poor stars ([Fe/H]$\leq -4.0$). The 80th percentile for the metallicity cumulative distribution function of the observed sample is [Fe/H]$= -2.04$. The sample also includes 68 Carbon-Enhanced Metal-Poor (CEMP) stars. Based on the calculated metallicities, further S-PLUS, color cuts are proposed, which can increase the fractions of stars with [Fe/H]$\leq -1.0$ and $\leq -2.0$ to $98\%$ and $88\%$, respectively. Such high success rates enable targeted high-resolution spectroscopic follow-up efforts, as well as provide selection criteria for fiber-fed multiplex spectroscopic surveys.