Background. Drug-resistant tuberculosis (TB) is an important threat in industrialized countries, but low information is known from Southern Italy. Here, we present the results of a retrospective study on TB cases diagnosed in 2008-2013 in Naples, the biggest city in the South of Italy. Methods. Six hundred ninety Mycobacterium tuberculosis strains were isolated at the Ospedali dei Colli of Naples, and resistance to first-line and second-line drugs was determined. Results. Multidrug-resistant (MDR) TB increased from 2008 to 2013, with most strains being isolated from migrants arriving from 41 countries. Overall MDR-TB rate was 4.5%: Italian-born persons, 2.2%; Romania, 7.5%; Former Soviet Union countries (Ukraine, Russia, Armenia, Georgia), 22.4%; all other foreign countries, 2.0%. Resistance of MDR strains to second-line drugs was high against kanamycin, ofloxacin, capreomycin. Conclusions. MDR-TB increased in 2008-13 and was mostly observed in migrants, indicating the need to intensify diagnosis and treatment of these populations in Naples.