| Objective:To understand the clinical status of clofazimine resistant gene of mycobacterium tuberculosis,and to provide a laboratory basis for the development of drug-resistant tuberculosis treatment programs containing clofazimine.Methods:Collecting clinical sputum specimens of outpatients and hospitalized tuberculosis patients in our hospital,conducting smear acid-fast staining,Mycobacterium tuberculosis culture,strain identification and drug sensitivity test(including first-line and second-line anti-tuberculosis drugs),screening clinical strains of Mycobacterium tuberculosis,extracting The Mycobacterium tuberculosis DNA,then expanding full length of the Clofazimine-resistant gene(Rv0678,Rv2535 c,Rv1979c)by polymerase chain reaction,and using the gene sequencing to obtain the M.tuberculosis resistance genotype results.Finally,the results of gene sequencing and M.tuberculosis standard strain H37 Rv gene sequence were compared with the US National Center for Biotechnology Information to further analyze the results.Results:A total of 121 clinical strains of Mycobacterium tuberculosis(60 cases of sensitive Mycobacterium tuberculosis,5 cases of monoresistance Mycobacterium tuberculosis,5 cases of polydrug resistance Mycobacterium tuberculosis and 51 cases of multi-drug resistant Mycobacterium tuberculosis)were obtained,and three clofazimine-resistant genes: Rv2535 c,Rv1979c and Rv0678 were detected.three clofazimine-resistant genes mutations were not detected in the susceptible strains,and the mutation rate was 0%(0/60).three clofazimine-resistant genes were not detected in monoresistance and polydrug resistance strains,and the mutation rate was 0%(0/5).in the multi-drug resistant Mycobacterium tuberculosis,Two mutations of Rv0678(GCC153GTC,ATG146AGG)were detected,and the mutation rate was 3.9%(2/51);Two mutations of Rv2535c(ACC56GCC,TTG121GTG)were detected,and the mutation rate was 3.9%(2/51);no Rv1979 c mutation was detected.The total gene mutation rate of the clofazimine-resistant gene Rv0678 and Rv2535 c of Mycobacterium tuberculosis clinical strains was 1.7%(2/121).Conclusion: There are mutations of clofazimine resistance gene in drug-resistant t uberculosis,and the risk of clofazimine resistance exists.Therefore,mutations in dr ug resistance genes should be considered in the development of anti-tuberculosis ch emotherapy regimens containing clofazimine.In order to reduce the risk of drug res istance,it is necessary to perform the clofazimine susceptibility test and drug resist ance gene detection before administration,At the same time,The discovery of the f our new mutations of clofazimine-resistance gene lay a laboratory foundation for f urther study of the resistance mechanism of clofazimine. |
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