摘要
目的应用基因芯片方法检测结核分枝杆菌(Mycobacterium tuberculosis,Mtb)对利福平和异烟肼的耐受性,评价其临床应用价值。方法应用聚合酶链反应(polymerase chain reaction,PCR)扩增-基因芯片杂交的方法检测经常规药敏实验证实的30株Mtb利福平和异烟肼敏感株和50株耐利福平和异烟肼分离株的rpoB基因及katG和inhA基因突变,同时以PCR-直接测序法为对照。结果应用PCR-基因芯片与基因测序方法检测30株Mtb利福平敏感株rpoB基因和异烟肼敏感株katG基因和inhA基因均为野生型。50株Mtb利福平耐药株中,PCR-基因芯片与基因测序分析3株rpoB基因均为野生型,41株均为突变型;6株PCR-基因芯片与基因测序结果不一致。50株Mtb异烟肼耐药株中,PCR-基因芯片与基因测序分析16株katG基因和30株inhA基因均为野生型,31株katG基因均为315位密码子突变,7株inhA基因均为-15位突变型,其中2株为katG和inhA双重突变;3株katG和13株inhAPCR-基因芯片与基因测序结果不一致。结论应用PCR-基因芯片方法可快速、有效地检出大多数Mtb耐多药分离株,指导临床用药。
Objective To detect Mycobacterium tuberculosis（Mtb）resistance to rifampicin（RIF） and isoniazid（INH） by gene chip,and to evaluate their clinical values. Methods Thirty RIF-and INH-susceptible and 50 RIF-and INH-resistant clinical isolates in Mtb were analyzed the mutations of rpoB、katG and inhA genes by（PCR）-gene chip and DNA sequencing. Results The rpoB、katG and inhA genes from 30 RIF-and INH-susceptible isolates of Mtb were all wild types by PCR-gene chip and DNA sequencing.Of 50 RIF-resistant clinical isolates,3 had wild types of ropB genes,and 41 had ropB mutations by PCR-gene chip and DNA sequencing.Six had inconsistent results between PCR-gene chip and DNA sequencing.Of 50 INH-resistant clinical isolates,16 had wild types of katG genes,and 31 had katG mutations on codon 315;30 had wild types of inhA genes,and 7 had inhA mutations at position-15.KatG results of 3 isolates and inhA results of 13 isolates were inconsistent between PCR-gene chip and DNA sequencing. Conclusion PCR-gene chip might be a rapid and effective method for the detection of Mtb multi-drug-resistant isolates.
出处
《中国防痨杂志》
CAS
2011年第10期680-685,共6页
Chinese Journal of Antituberculosis
基金
北京市科技重大专项重点项目(Z09050700940904)
"十一五"国家重大科技专项(2008ZX10003-001)
关键词
分枝杆菌
结核
利福平
异烟肼
抗药性
细菌
寡核苷酸序列分析
Mycobacterium tuberculosis
Rifampicin
Isoniazid
Drug resistance
bacterial
Oligonucletide array sequence analysis
