结核分枝杆菌外排泵基因表达与耐药相关性研究

Association between ATP-binding cassette efflux pump gene drrA,drrB,pstB and Rv1273c expression and drug resistance of mycobacterium tuberculosis

目的探讨结核分枝杆菌(mycobacterium tuberculosis,MTB)ATP结合盒外排泵基因的表达与不同耐药表型的关系,为耐药结核病的有效防控提供科学依据。方法收集宁波市疾病预防控制中心的99例结核分枝杆菌作为研究对象。提取菌株RNA,应用RT-PCR方法检测ATP结合盒外排泵基因drrA、drrB、pstB及Rv1273c的表达量,采用Mann-Whitney u检验分析外排泵基因表达在不同耐药表型菌株中的差异。结果 pstB及Rv1273c基因表达量在耐利福平组(2.449、0.765)高于利福平敏感组(1.949、0.498),差异有统计学意义(Z=1.970、P=0.049,Z=2.330、P=0.020)。耐多药组drrB、pstB及Rv1273c基因表达量(2.481、2.702、0.884),高于非耐多药组(2.224、1.884、0.495),差异均有统计学意义(Z=2.195、2.892、2.741,P均<0.05)。异烟肼耐药组和敏感组中Rv1273c的高表达率均差异有统计学意义(P<0.05),利福平耐药组和敏感组中drrA、drrB、pstB及Rv1273c的高表达率均差异有统计学意义(P<0.05),乙胺丁醇耐药组和敏感组中pstB及Rv1273c的高表达率均差异有统计学意义(P<0.05)。结论 ATP结合盒外排泵基因pstB及Rv1273c的表达与结核分枝杆菌对利福平的耐药有关。

Objective To explore the association between ATP-binding cassette efflux pump gene drrA,drrB,pstB and Rv1273 c expression and drug resistance of mycobacterium tuberculosis.Methods A total of 99 mycobacterium tuberculosis isolates including 66 which were resistant to one or more drugs as isoniazid,rifampicin,streptomycin,ethambutol and 33 isolates which were sensitive to the above four drugs collected in Ningbo CDC were retrieved for the study.RNA was extracted and ATP-binding cassette efflux pump gene drrA,drrB,pstB and Rv1273 c were amplified by RT-PCR.Data were analyzed by Mann-Whitney u test.Results pstB and Rv1273 c gene in isolates resistant to rifampicin and drrB,pstB,Rv1273 c gene in isolates with multi-drug resistance were all up-regulated compared to those sensitive to rifampicin and those without,the differences were statistically significant(P<0.05).The over-expression rates of Rv1273 c gene between isoniazid-resistant and isoniazid-susceptible isolates,drrA,drrB,pstB and Rv1273 c genes between rifampicin-resistant and rifampicin-susceptible isolates and pstB,Rv1273 c genes between ethambutol-resistant and ethambutol-susceptible isolates were all significantly different(P<0.05).Conclusions The expression of ATP-binding cassette efflux pump gene pstB and Rv1273 c are associated with the resistance of mycobacterium tuberculosis.To rifampicin.