Genotyping Of Resistant Ofloxacin M.Tuberculosis Clinical Isolates Strains And Effect Of Pump Inhibitor Drug Sensitivity

Objiective:The purpose of this study is evaluated three kinds of efflux pump inhibitors reserpine, verapamil and carbonyl cyanogen chlorobenzene hydrazone, They resistance to effect of ofloxacin clinical separation strains,and by analyzing thses results of MIC medicine sensitivity,Determine the effect of effux pumps in resistance.then carry through genotyped and the results were analyzed to provide avidence for the clinical diagnosis.Methods:Thirty-two mycobacterium tuberculosis clinical separation strains (twenty-eight ofloxacin strains resistant strains and four ofloxacin sensitive strains), Through resazurin reagent experimental observation existence of three kinds of efflux pump inhibitors (reserpine, verapamil and carbonyl cyanogen chlorobenzene hydrazone),its the MIC is worth change, And in contrast with the control group (not add pump inhibitors group).Then make ues of the Spoligotyping classification method and the VNTR classification point to this mycobacterium tuberculosis clinical separation srains gene type, And by the NatureEdge software on data clustering analysis, observe the relationship between them.Results:By the results of MIC ofloxacin drug sensitive experiment, show that thirty-two strains mycobacterium tuberculosis clinical separation strain, Four strains of mycobacterium tuberculosis for ofloxacin sensitive type and twenty-eight strains have different degrees of drug-resistant strains. Add to three efflux pump inhibitors results indicate that four ofloxacin sensitive strains MIC value are not change significantly, and twenty-eight strains resistant clinical separation strains is change apparently.Reserpine group average is thirty-two fold decrease, verapamil and carbonyl cyanogen chlorobenzene hydrazone groups are sixteen fold decrease, These two groups found a bacteria MIC values did not change significantly for further study. And by Spoligotyping and VNTR this two genotype methods carry through thirty-two clinical separation strains genotyping, The results found that twenty-seven strains bacteria for Beijing genetype, one strains bacteria for T2type and four strains isn’t record in the Spoligotyping database.Results of VNTR and the type of Spoligotyping results are consistent. And found that drug-resistant strains with Beijing gene type is major, four special genetype for drug-resistant strains, and the relative relation is very close, they have certain representativeness.Conclusions:by this MIC medicine sensitivity experiments show that,compare with traditional methods,it simple operation, clear observation, and simple equipment, low cost, and it is a certain use prospects. For these mycobacterium tuberculosis clinical separation strains gene classification, we found that most of the drug-resistant genotyping for Beijing genetype.