ObjectiveTo determine the extent of drug resistance in multidrug-resistant tuberculosis (MDR-TB) cases, we conducted a retrospective, population-based analysis using drug susceptibility testing (DST) results of MDR Mycobacterium tuberculosis complex isolates obtained from 2007–2014 in Taiwan.MethodsM. tuberculosis isolates collected from 1,331 MDR-TB cases were included in this survey. Treatment histories, age, sex, chest radiograph and bacteriological results of patients were analyzed. Standard DST was performed to assess resistance to the following drugs: isoniazid (INH), rifampicin (RIF), streptomycin (SM), ethambutol (EMB), amikacin (AM), kanamycin (KM), capreomycin (CAP), ofloxacin (OFX), moxifloxacin (MOX), levofloxacin (LVX), gatifloxacin (GAT), para-aminosalicylate (PAS), ethionamide (EA), and pyrazinamide (PZA). The Cochran-Armitage trend test was used for statistical analysis.ResultsWe observed a significant increasing trend in portion of new MDR-TB cases, from 59.5% to 80.2% (p < 0.0001), and significant decreasing trend of portion in the 15-44-year-old age group (p < 0.05). Of the MDR M. tuberculosis isolates tested, 6.2% were resistant to AM, 8.6% were resistant to KM, 4.6% were resistant to CAP, 19.5% were resistant to OFX, 17.1% were resistant to MOX, 16.0% were resistant to LVX, 5.8% were resistant to GAT, 9.5% were resistant to PAS, 28.5% were resistant to EA and 33.3% were resistant to PZA. Fifty (3.8%) extensively drug-resistant TB cases were identified. No significant differences were found in drug resistance frequencies between new and previously treated MDR cases. However, we observed significant decreases in the rates of AM resistance (p < 0.05), OFX resistance (p < 0.00001), PAS resistance (p < 0.00001), EA resistance (p < 0.05) and PZA resistance (p < 0.05). Moreover, younger age groups had higher rates of resistance to fluoroquinolones.ConclusionA policy implemented in 2007 to restrict the prescription of fluoroquinolones was shown to be effective. Our survey revealed a decreasing trend of resistance to PZA, OFX and AM, which suggests the feasibility of adopting a short-course regimen and demonstrates the effectiveness of our management program for MDR-TB.