基因芯片快速鉴定分枝杆菌菌种方法的建立  

Establishment of a rapid and accurate gene microarray for identifying Mycobacterium species

作　　者：唐曙明[1] 李爱敏[1] 陈海霞[1] 杨自华[1] 

机构地区：[1]深圳市人民医院检验科,广东深圳518000

出　　处：《重庆医学》2015年第11期1516-1518,共3页Chongqing medicine

基　　金：深圳市科技局资助项目(201102151)

摘　　要：目的利用基因芯片技术,建立一种能快速、准确鉴定分枝杆菌菌种的方法,并验证其临床应用价值。方法根据23种分枝杆菌基因序列设计特异性探针并制作基因芯片,通过PCR-反向点杂交鉴定23种分枝杆菌标准菌株、9种非分枝杆菌和103株分枝杆菌临床分离株的菌种。结果应用基因芯片技术检测23种分枝杆菌标准菌株和9种非分枝杆菌菌株,特异性为100%。103株临床分离株经鉴定87株为结核杆菌复合群(MTC);16株为非结核分枝杆菌(NTM),其中脓肿分枝杆菌5株、胞内分枝杆菌3株、鸟分枝杆菌3株、偶发分枝杆菌2株,以及堪萨斯分枝杆菌、海分枝杆菌、戈登分枝杆菌各1株。103株临床分离株鉴定结果与测序法完全一致,该方法最低检出限为103 copy/mL。结论采用基因芯片技术能快速鉴定分枝杆菌菌种,并区分MTC和NTM,具有简便快速及准确性、特异性、灵敏度高的优点。Objective To establish a rapid and accurate method for the identification of Mycobacterium species by the gene microarray and to verify its clinical application value .Methods According to the gene sequence of 23 species of Mycobacteria ,the specific probes were designed and the gene chips were prepared .23 Mycobacterial standard strains ,9 non‐mycobacterial strains ,103 clinically isolated mycobacterial strains were detected by PCR‐based reverse blot hybridization assay in the gene chip .Results 23 mycobacterial standard strains ,9 non‐mycobacterial strains were detected by gene chip ,the results showed that the specificity was 100% .Of 103 mycobacterial clinically isolated strains ,87 strains were identified as Mycobacterium tuberculosis compounds （MTC） and 16 strains as non‐tuberculosis mycobacteria （NTM ） including 5 strains of M .abscessus ,3 strains of M .intracellulare ,3 strains of M .avium ,2 strains of M .fortuitum ,1 strain of M .kansas ,1 strain of M .marinum and 1 strain of M .gordonae .The identification results of 103 clinically isolated strains were completely consistent with the sequencing results .The lowest detection limit by this method was 103 copies/mL .Conclusion The gene microarray technique for rapidly identifying Mycobacteria and differentiate MTC and NTM has the advantages of simpleness ,rapidness ,high accuracy ,high specificity and high sensitivity .

关 键 词：分枝杆菌属 菌种鉴定 寡核苷酸序列分析 

分 类 号：R446.5[医药卫生—诊断学]