摘要
目的探讨基因芯片检测系统在分枝杆菌菌种鉴定及耐药基因检测中的临床应用价值。方法应用基因芯片技术对疑似结核病和非结核分枝杆菌病患者的痰液、尿液、胸腹腔积液、脑脊液和穿刺液等标本进行核酸检测;并对分枝杆菌阳性样本进行利福平及异烟肼的耐药基因检测。结果 91例标本中共检出结核分枝杆菌复合群16例,非结核分枝杆菌2例(戈登分枝杆菌1例、偶然分枝杆菌1例),分枝杆菌检出总阳性率为19.8%(18/91),其中结核分枝杆菌占17.6%(16/91),非结核分枝杆菌占2.2%(2/91);不同标本类型检出阳性率有差别,由高到低依次为穿刺脓液(33.3%)、痰液(24.0%)、胸腹腔积液(18.8%)、脑脊液(16.7%)和尿液(15.8%)。有57例患者同时进行了基因芯片法和涂片抗酸染色法检测,其中芯片法有16例阳性,阳性率为28.1%(16/57),涂片法有3例阳性,阳性率为5.3%(3/57),两种方法比较,差异有统计学意义(P<0.01)。18例分枝杆菌阳性标本进一步进行耐药基因检测,有4例发生耐药基因突变,其中2例(1例戈登分枝杆菌)为利福平耐药相关基因,ropB基因526位点发生C→G突变,1例为ropB基因531位点发生C→T突变,还有1例为异烟肼耐药相关基因,inhA基因启动子-15位点发生C→T突变;没有检测到多重耐药株。结论基因芯片技术适用于不同标本类型的分枝杆菌菌种鉴定及耐药基因检测,操作简便快速,灵敏度高,特异性好。
Objective To investigate the diagnostic value of gene chip in identification of mycobacterium and detection of drug-resistant genes .Methods Gene chip technology was used in DNA detection of sputum ,urine ,pleu-ral effusion ,cerebrospinal fluid ,and puncture fluid samples from suspected tuberculosis and nontuberculous mycobac-teria patients ,and positive samples of mycobacterium were detected for drug-resistant genes of mycobacterium to ri-fampin and isoniazid further .Results Among 91 samples detected ,16 cases of mycobacterium tuberculosis complex , 2 cases of nontuberculous mycobacteria （1 case of Gordon′s bacillus ,1 case of mycobacterium fortuitum ） ,the positive rate of mycobacterium was 19 .8% （18/91） ,which the mycobacterium tuberculosis accounted for 17 .6% （16/91）;Mycobacterium non tuberculosis accounted for 2 .2% （2/91） .The positive rate of various types of samples were dif-ferent ,which puncture pus ,sputum ,pleural effusion ,cerebrospinal fluid and urine accounted for 33 .3% ,24% , 18 .8% ,16 .7% and 15 .8% .57 patients were detected by the gene chip method and mycobacterium smear acid-fast staining at the time ,the chip was positive in 16 cases ,the positive rate was 28 .1% （16/57） ,smear was positive in 3 cases ,the positive rate was 5 .3% （3/57） ,there was significant difference between the two methods （P〈0 .01） .18 ca-ses of Mycobacterium positive samples were detected for drug-resistant genes further ,4 cases showed drug-resistant genes mutation ,2 cases（1 case Gordon mycobacteria） were ropB gene 526 C → G mutation ,1 case was ropB gene 531 C → T mutation ,1 case was InhA gene promoter-15 C → T mutation .Multiple resistant strains were not detected . Conclusion The gene chip technology can be used for mycobacterial species identification and drug-resistant genes detection In different types of samples ,which is simple and rapid in procedure and have high sensitivity and good spe-cificity .
出处
《检验医学与临床》
CAS
2014年第12期1595-1598,共4页
Laboratory Medicine and Clinic
基金
辽宁省社会发展基金资助项目(2011225019)
关键词
基因芯片
分枝杆菌
鉴定
耐药
gene chip
mycobacterium tuberculosis
identification
drug resistance
