CAJ | 학술논문

目的采用变性高效液相色谱(DHPLC)法和测序法来分析结核分枝杆菌链霉素耐药基因rpsL和rrs基因突变,从而检测是否对链霉素耐药.方法 215株结核分枝杆菌临床分离株(经常规方法证实,115株为链霉素耐药,100株为敏感),测定链霉素最小抑菌浓度(MIC),同时采用DHPLC和测序法检测rpsL和rrs基因突变.结果 85.2%(98/115)的链霉素耐药株携有rpsL和(或)rrs基因突变,其中rpsL基因突变占大多数(76.5%,88/115).对98株带有基因突变的菌株的分析表明,rpsL,rrs基因的突变类型与MIC之间未见密切关系.100株敏感株未检出基因突变.DHPLC法与测序法结果完全一致.结论本工作建立的DHPLC法可以认为是结核分枝杆菌链霉素耐药分析的一种有效的工具.
목적 채용변성고효액상색보(DHPLC)법화측서법래분석결핵분지간균련매소내약기인rpsL화rrs기인돌변,종이검측시부대련매소내약.방법 215주결핵분지간균림상분리주(경상규방법증실,115주위련매소내약,100주위민감),측정련매소최소억균농도(MIC),동시채용DHPLC화측서법검측rpsL화rrs기인돌변.결과 85.2%(98/115)적련매소내약주휴유rpsL화(혹)rrs기인돌변,기중rpsL기인돌변점대다수(76.5%,88/115).대98주대유기인돌변적균주적분석표명,rpsL,rrs기인적돌변류형여MIC지간미견밀절관계.100주민감주미검출기인돌변.DHPLC법여측서법결과완전일치.결론 본공작건립적DHPLC법가이인위시결핵분지간균련매소내약분석적일충유효적공구.
Objective To determine the rpsL and rrs gene mutation in Mycobacterium tuberculosis (M. tuberculosis)and compare the consistency between the results of denaturing high-performance liquid chromatography(DHPLC)and those of DNA sequencing. Methods The values of streptomycin minimum inhibitory concentration(MIC)against 215 M. tuberculosis clinical isolates, 115 being streptomycinresistant and 100 being susceptible by a routine proportional method, were tested by DHPLC. DNA sequencing was conducted to detect the rpsL and rrs mutation. Results 98 of the 115 streptomycin-resistant isolates(85. 2%)harbored rpsL and/or rrs mutation, 76. 5%of which being rpsL mutation(88/115). There was no significant correlation between the MIC values and mutation types. No mutation was found in all the susceptible isolates. There was a complete consistency between the DHPLC results and those of DNA sequencing. Conclusion DHPLC can be regarded as a useful and powerful tool to detect the streptomycin resistance detection in M. tuberculosis.