Antimicrobial resistance mechanisms among clinically relevant gram-positive microorganisms can be demonstrated using phenotypic tests that enable the interpretation of underlying mechanisms responsible for the in vitro resistance. The reporting of these mechanisms, either inferred or demonstrated, helps in the adjustment of clinical treatments and the epidemiological follow up of resistance traits. In the present work, phenotypic tests for detection of antimicrobial resistance mechanisms involving the most frequent antimicrobial families used against Staphylococcus spp., Enterococcus spp. and Streptococcus pneumoniae are analysed. In the case of Staphylococcus, phenotypic tests to reveal the mechanisms of resistance against beta-lactams, macrolides, lincosamides and streptogramin B (MLS(B)), as well as intermediate susceptibility to glycopeptides, resistance to aminoglycosides, mupirocin and linezolid are reviewed. Tests to detect glycopeptide resistance and high-level aminoglycoside resistance among enterococci are analysed. Detection of penicillin resistance, as well as diminished susceptibility to third generation cephalosporins, together with diminished susceptibility or even resistance to fluoroquinolones is also detailed.