基因芯片技术检测结核分枝杆菌利福平和异烟肼耐药性临床应用评价  

Evaluation for clinical application effect of gene chip for detection of rifampin and isoniazid resistance in Mycobacterium tuberculosis

作　　者：许榕青[1] 李丹[1] 林银霞 黄明翔[1] 陈新朝[1] 

机构地区：[1]福建省福州肺科医院检验科,福建医科大学临床教学医院,福州350008

出　　处：《中国人兽共患病学报》2017年第1期43-48,共6页Chinese Journal of Zoonoses

基　　金：福建省卫生厅青年科研课题项目(No.2013-2-77)~~

摘　　要：目的评价基因芯片技术检测结核分枝杆菌利福平和异烟肼耐药性的临床应用效果。方法利用基因芯片技术对涂阳肺结核患者的痰标本和临床分离株进行利福平和异烟肼耐药基因位点检测,比较痰标本与菌株检测结果的差异;并以BACTEC MGIT 960药敏试验结果为对照评价基因芯片的检测性能。另外,通过对999株鉴定为结核分枝杆菌的临床分离菌株进行rpoB、katG和inhA基因耐药相关区域扩增产物测序,以验证基因芯片对核酸序列检测的准确性。结果在涂阳的1 108例标本中有100例经基因芯片菌种鉴定为非结核分枝杆菌。其余的1 008例痰标本基因芯片结果与BACTEC MGIT960培养阳性的菌株基因芯片结果比较仅有9例结果不一致,符合率为99.1%。以BACTEC MGIT960培养法药敏结果为对照,999株菌株基因芯片法检测利福平耐药性的符合率为98.1%,异烟肼的耐药性的符合率为94.5%。与DNA测序法相比,基因芯片法检测利福平和异烟肼耐药性的准确率分别为99.6%和99.8%。结论基因芯片技术能够快速、准确地进行结核分枝杆菌利福平和异烟肼的耐药性检测,并能直接用于痰标本检测。We evaluated clinical application effect of gene chip for detection of rifampin （RFP） and isoniazid （INH） resistance in Mycobacterium tuberculosis （MTB）. Rifampin and isoniazid drug-resistance gene loci were detected by gene chip with sputum specimens from smear-positive tuberculosis patients and clinical strains, comparing the results of detection. BACTEC MGIT 960 drug susceptibility test results were used as control to evaluate the detection performance of gene chip. The sequences of the polymerase chain reaction products of the rpoB, katG and inhA genes from 999 strains identified as Mycobacterium tuberculosis were determined to confirm the mutations by DNA sequencing. Results showed that 100 cases were identified as nontuberculous mycobacteria by gene chip in the 1 108 cases of smear-positive samples. Among the rest 1 008 samples, there were only 9 cases of microarray results different from BACTEC MGIT960 culture-positive strains, achieving the coincidences of 99.1%. Compared with BACTEC MGIT 960 drug susceptibility test results, the gene chip method displayed a concordance of 98.1% and 94.5% for RFP and INH respectively in the 999 strains. Compared with the DNA sequencing method, the accuracy of gene chip method was 99.6% for rifampin resistance and 99.8% for isoniazid resistance. It＇s concluded that the gene chip technology can quickly and accurately detect rifampin and isoniazid resistance in MTB and can be used directly for the detection of sputum samples.

关 键 词：结核分枝杆菌 利福平 异烟肼 耐药性 基因芯片 

分 类 号：R378.91[医药卫生—病原生物学]