结核菌WhiB7,drrA,drrB基因表达与其抗结核药物的耐受关系研究  

Research of the relationship between the expression of mycobacterrium tuberculosis WhIB7,drrA,drrB genes and anti-tuberculosis drug tolerance

作　　者：王星 史建中[2] 刘镇[2] 陈向东 李海 谢兰品[4] 吕翠环[4] 陈素丽[4] 刘金录[5] 邱景富[5] 

机构地区：[1]张家口市肺科医院,河北怀来县075441 [2]张家口市疾病预防控制中心,河北张家口075000 [3]张家口市第六医院,河北张家口075000 [4]河北省胸科医院,河北石家庄050000 [5]河北北方学院,河北张家口075000

出　　处：《临床肺科杂志》2016年第12期2178-2183,共6页Journal of Clinical Pulmonary Medicine

基　　金：河北省科技计划项目(No 142777100D)

摘　　要：目的探讨结核分枝杆菌耐药性与WhIB7,drrA,drrB基因表达的关系。方法通过设计并合成结核分枝杆菌的Whi B7,drrA,drrB基因引物,对耐药的结核分枝杆菌利用逆转录得到三个基因的c DNA。并对c DNA进行PCR扩增,通过优化PCR参数,使PCR方法达到检测c DNA拷贝数的准确性。分析WhIB7,drrA,drrB基因表达在不同类型耐药菌株的表达程度。使用药物刺激观察耐药菌株Whi B7,drrA,drrB基因的变化。结果逆转录荧光定量PCR可以对WhIB7,drrA,drrB基因很好的定量。对不同种耐药菌株进行WhIB7,drrA,drrB基因检测,阳性率分别为:单利福平耐药菌株85.0%(17/20)、35.0%(7/20)、30.0%(6/20),单异烟肼耐药菌株37.5%(12/32)、87.5%(28/32)、81.3%(26/32),利福平和异烟肼均耐药的菌株87.5%(14/16)、75.0%(12/16)、81.3%(13/16),含利福平异烟肼的多重耐药菌株88.2%(30/34)、82.4%(28/34)、85.3%(29/34),药物敏感菌株12.4%(12/97)、8.2%(8/97)、9.3%(9/97)。耐药菌株WhIB7,drrA,drrB基因阳性率高于药物敏感菌株。WhIB7基因与利福平耐药关系紧密,drrA和drrB基因与异烟肼耐药关系紧密。但不同的药物对耐药菌基因表达的诱导能力不同。结论 Whi B7,drrA,drrB基因的逆转录荧光定量PCR方法可以作为耐药结核分枝杆菌筛查的重要参考,为研究结核分枝杆菌日后基因表达与耐药机理研究奠定基础。Objective To explore the relationship between the expression of mycobacterrium tuberculosis WhIB7, drrA, drrB genes and anti-tuberculosis drug tolerance. Methods We designed and synthetized the primers of gene WhiB7, drrA and drrB, and cDNAs of the three genes＇ mRNA were obtained by reverse transcription. Then the cDNAs were augmented by polimerase chain reaction. Parameters were optimized to achieve the best degree of accuracy of cDNA＇ s copy number. And then we researched the degree of genes＇ expression in drug-fast bacterial strains. It used medicine inducing to observe the changes of hiBT, drrA and drrB genes. Results Reverse transcription fluores- cent quantitation PCR had good determination on WhIB7, drrA and drrB genes. The positive rate of RFP-resistant strain for WhlB7, drrA and drrB genes was 85.0% （ 17/20）, 35.0% （7/20） and 30. 0% （6/20） respectively, INH-resistant strain for 37.5% （ 12/32）, 87. 5% （28/32） and 81.3% （26/32）, RFP＆INH-resistant strain for 87. 5% （ 14/16）, 75.0% （12/16） and 81.3% （ 13/16）, multiple drug-fast bacteria for 88.2% （30/34）, 82.4% （28/34） and 85.3% （29/34）, and drug susceptibility strain for 12.4%, 8. 2% and 9. 3%. Generally, the positive rate of WhIB7, drrA and drrB expression in drug-fast bacterial was more than drug-susceptibility bacterial. WhlB7 gene had more close-knit relationship with RFP drug-fast, and drrA and drrB gene had more close-knit relationship with INH drug-fast. Moreover, different drugs had different ability of inducing drug-fast degree. Conclusion It is an important reference to screen drug-fast mycobacterium tuberculosis using RT-PCR of WhIB7, drrA and drrB genes, which may establish foundation to find out the mechanism of expression and drug-fast in mycobacterium tuberculosis.

关 键 词：利福平 异烟肼 WhIB7 drrA drrB 

分 类 号：R440[医药卫生—诊断学] R52[医药卫生—临床医学]