SURVEYOR酶法检测结核分枝杆菌乙胺丁醇耐药基因突变的探讨  

Drug-resistance gene mutation analysis of mycobacterium tuberculosis by SURVEYOR nuclease

作　　者：张霞[1] 赵德华[1] 秦殊[2] 石瑞如[2] 张国龙 

机构地区：[1]郑州大学第三附属医院,郑州450052 [2]河南省胸科医院 [3]河南省公共卫生医疗中心

出　　处：《医药论坛杂志》2012年第12期45-46,共2页Journal of Medical Forum

摘　　要：目的探讨SURVEYOR酶法在结核分枝杆菌耐药基因突变检测中的应用。方法结核分枝杆菌临床分离株60株,经测序证实有embB基因突变的乙胺丁醇耐药株45株,无基因突变的15株,采用SURVEYOR酶法进行检测。经PCR扩增,杂交形成异源双链,SURVEYOR酶切,通过聚丙烯酰胺凝胶电泳的带形来判断有无突变存在。结果 45株耐药株均检出了存在突变,15株敏感株均无突变发现。SURVEYOR酶法与测序法的结果完全相符。结论 SURVEYOR酶法检测结核分枝杆菌乙胺丁醇耐药基因突变简便、稳定,灵敏度和特异性高,是结核分枝杆菌耐药基因检测较好方法。Objective To explore the application of SURVEYOR Nuclease in drug-resistance gene mutation analysis of mycobacterium tuberculosis.Methods SURVEYOR Nuclease were used to analyze embB gene mutation of 60 with clinical isolates,among which 45 were EMB-resistant and found embB gene mutation by sequencing and 15 EMB-susceptible isolates without embB mutation.First PCR amplification,then hybridized with products of H37Rv to form heteroduplex,cleaved by SURVEYOR Nuclease,at last,results were shown by polyacrylamide gel electrophoresis and mutation was judged according to electrophoresis profile.Results Totally 45 EMB-resistant isolates were all found harbouring embB gene mutation while those 15 EMB-susceptible isolates no mutation was found.These results from SURVEYOR Nuclease method is completely parallel to those of sequencing.Conclusions Heteroduplex analysis by SURVEYOR Nuclease may become a useful method for drug-resistance gene mutation analysis of mycobacterium tuberculosis since its simplicity,stability,low-cost,high sensitivity,specificity and no need for special equipment.

关 键 词：SURVEYOR酶法 结核分枝杆菌 MDR 

分 类 号：R446.5[医药卫生—诊断学]