Following the World Health Organization's endorsement of the Xpert® MTB/RIF assay, which rapidly and simultaneously diagnoses tuberculosis (TB) and detects resistance to rifampin (RMP), the question arises to what extent RMP resistance is an adequate marker for multidrug-resistant TB (MDR-TB). A retrospective analysis of data from >81 countries and subnational settings demonstrated that >40% of RMP-resistant isolates from new TB cases did not display resistance to isoniazid (INH) in settings with relatively low MDR-TB prevalence (one third of all countries and subnational settings). Results indicated the need for INH susceptibility testing in addition to RMP susceptibility testing.