结核分枝杆菌耐药分析及基因检测研究

An Investigation on Multidrug Resistant status and gene detection of Mycobacterium Tuberculosis in Changchun City

目的了解长春市结核分支杆菌菌株的耐药状况,建立适合我国国情的结核分支杆菌耐药株的快速检测手段。方法采用普通改良罗氏培养,对2004年7月-2007年12月间分离培养的结核分支杆菌进行菌株鉴定及药物敏感实验,并利用PCR反向斑点杂交技术(RDB)和测序方法对rpoBr、psL、KatG和embB基因进行检测。结果 625株结核分支杆菌中,其耐药比例为32.96%。耐药顺位次序为INH 18.17%、RIF 17.26%S、M 10.74%、EMB 7.31%。复治患者的耐药率高于初治患者。在随机抽取的菌株中,RDB检出的rpoBr、psL、KatG和embB基因突变率分别为88.9%、83.3%、91.6%和86.1%,和测序结果的符合率分别为87.9%,87.9%、91.4%和89.1%。结论长春市结核病患者总耐药比例高于我国结核病总耐药率,耐药状况具有本市的耐药特点,耐药的产生与菌株基因突变有较为密切的关系。结核病人耐药情况比较严重,需要更严格的执行结核病控制策略,防止耐药病人及耐药菌株的产生和传播。

Objective To find out the muhidrug resistant situation of Mycobacterium Tuberculosis in Changchun City in order to establish fast diction method according to the status in our country.Methods By modified Roche media,strains isolated between Jul 2004 to Dec 2007 were identified and the drug resistance was tested.PCR based reverse dot blot（RDB） and sequencing method were also used to analyze rpoB,rpsL,KatG and embB gene.Results The total multidrug resistant ratio of tuberculosis in Jilin Province is 32.96% among all the 625 strains;The drug resistance ratio is as follow：INH 18.17%、RIF 17.26%、SM 10.74%、EMB 7.31%.The ratio of retreated patients is higher than first treated ones.Among stochasticly selected strains,the RDB tested mutation rate of rpoB、rpsL,KatG and embB gene are 88.9%,83.3%,91.6% and 86.1% and compared rate with sequencing are 87.9%,87.9%,91.4% and 89.1%.Conclusion The drug resistant ratio in Changchun City is higher than that in the country and the situation has its own feature.The appearance of drug resistance has something to do with gene mutation.The situation is serious and need more restrict disease control strategy to avoid spread.