新基因MTB73与儿童结核病耐药相关性的初步研究  

Preliminary study on the correlation of gene MTB73 with drug resistant tuberculosis in children

作　　者：苏薇[1] 黄延风[1] 郑改焕[1] 朱朝敏[1] 郭倩[1] 

机构地区：[1]重庆医科大学附属儿童医院感染科,儿童发育疾病研究教育部重点实验室,儿科学重庆市重点实验室,重庆400014

出　　处：《第三军医大学学报》2016年第18期2082-2086,共5页Journal of Third Military Medical University

基　　金：重庆市卫生和计划生育委员会医学科研计划基金(20142047)~~

摘　　要：目的通过检测儿童结核感染临床分离株的耐药表型及其MTB73基因的表达,探讨该基因与结核耐药性的关系。方法 110株儿童结核病临床分离株通过药敏实验分为耐药组和敏感组两个组,应用RT-PCR和基因测序的方法检测并验证结核分离株中新基因MTB73的表达,应用定量PCR检测基因MTB73在菌株中的表达量,比较耐药组菌株和敏感组菌株中MTB73基因阳性表达率和表达量的差异。结果 110例临床结核分离株样本中MTB73基因阳性表达率为11.82%(13株)。经比较,耐药结核菌中MTB73阳性表达率及相对表达量均明显高于敏感菌株(χ2=6.014,P=0.014;t’=8.075,P<0.001)。根据耐药种数不同,将耐药菌株分为耐单药(32株)、多耐药(12株)和耐多药(14株)3个亚组,其中耐多药组的MTB73阳性表达率(9株,64.29%)明显高于单耐药组(1株,3.13%)、多耐药组(1株,8.33%)及敏感组(2株,3.85%),差异有统计学意义(χ2=17.969,P<0.001;P=0.005;χ2=24.823,P<0.001)。单耐药组、多耐药组及敏感组3个组中MTB73基因阳性表达率两两之间比较差异无统计学意义(P>0.008)。结论 MTB73基因在耐药菌株尤其是耐多药菌株中高表达,提示新基因MTB73可能与儿童耐药结核病的耐药性相关。Objective To study the correlation of gene MTB73 to drug resistant tuberculosis （TB） in children by measuring MTB73 mRNA expression in isolated strains of Mycobacterium tuberculosis （M. tuberculosis） resistant isolates. Methods A total of 110 clinical isolates of children with TB were divided into resistant group and sensitive group by drug susceptibility test. Reverse transcriptase polymerase chain reaction （RT-PCR） and gene sequencing techniques were used to detect and verify MTB73 gene expression in the 110 clinical isolates. The expression level of MTB73 was quantified using SYBR Green I real-time PCR and the differences of the expression between these 2 groups were analyzed. Results Of 110 clinical isolates, 11.82% （13 isolates） were MTB73 expressed. The expression rate and level of MTB73 were significantly higher in drug resistant isolates than in drug sensitive isolates, with statistical significance （ Chi square = 6. 014,P = 0. 014 ;t＇ = 8. 075,P 〈 0. 001 ）. The drug resistant isolates were further divided into 3 subgroups ： single drug-resistance （n = 32）, polyresistanee （n = 12） and multidrug resistance （n = 14）. The expression rate of MTB73 in multidrug resistant isolates （9,64.29%）was significantly higher than in single drag-resistant group（ 1,3.13% ）, polyresistant group（ 1,8.33% ）and drug sensitive group（2,3.85% ）, with a statistically significant difference （ Chi square = 17. 969, P 〈 0. 001 ; P = 0. 005 ; Chi square = 24. 823, P 〈 0. 001 ）. Additionally, MTB73 expression was not significantly different among the single drug-resistant, polyresistant and drug sensitive groups （ P 〉 0. 008 ）. Conclusion Gene MTB73 is significantly upregulated in resistant isolates, especially in multidrug resistant isolates, suggesting that MTB73 may be associated with drug resistance of children TB.

关 键 词：结核分枝杆菌 新基因MTB73 聚合酶链反应 耐药性 

分 类 号：R372[医药卫生—病原生物学] R725.2[医药卫生—基础医学]