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Objective:To determine the patts of resistance to first line anti-tuberculosis(TB) drugs among a collection ofMycobacterium tuberculosis(MTB) isolates from5 provinces ofIran. Methods:A total of the6426 clinical specimens from patients suspected of activeTB were collected fromMarch2010 toJune2012.All specimens were subjected for microscopy and culture tests in theTB centers of studies provinces.Drug susceptibility testing to the first line anti-TB drugs for culture positiveMTB was performed onL?wenstein-Jensen(LJ) medium using proportion method.Results:Of6426 clinical specimens,261 were culture positive for mycobacteria, of which252 wereMTB and9 wereMOTT(mycobacteria other than tuberculosis). Of252MTB isolates,211(83.7%) were pan-susceptible and41(16.3%) were resistant to at least one drug.Resistance was most common to streptomycin,30 isolates(12.0%), followed by isoniazid,20 isolates(8.0%), rifampin,15 isolates(6.0%) and ethambutol,14 isolates(5.5%).Sixteen(6.3%)MTB isolates wereMDR.A clear evidence of heterogeneity amongst the5 provinces in the proportions with resistance to one or more drugs was observed [χ2 =12.209(4 degrees of freedom),P values =0.0159].Conclusions:The prevalence of drug resistance in this study area underscoring the need for further enforcement ofTB control strategies in theIran.Drug susceptibility testing for allTB cases to provide optimal treatment, establishing advanced diagnostic facilities for rapid detection ofMDR-TB and continuous monitoring of drug resistance are recommended for prevention and control of drug-resistantTB.