CAJ | 학술논문

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103copy／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。
목적：이용기인심편기술，건립일충능쾌속、준학감정분지간균균충적방법，병험증기림상응용개치。방법근거23충분지간균기인서렬설계특이성탐침병제작기인심편，통과PCR‐반향점잡교감정23충분지간균표준균주、9충비분지간균화103주분지간균림상분리주적균충。결과응용기인심편기술검측23충분지간균표준균주화9충비분지간균균주，특이성위100％。103주림상분리주경감정87주위결핵간균복합군（MTC）；16주위비결핵분지간균（NTM），기중농종분지간균5주、포내분지간균3주、조분지간균3주、우발분지간균2주，이급감살사분지간균、해분지간균、과등분지간균각1주。103주림상분리주감정결과여측서법완전일치，해방법최저검출한위103copy／mL。결론채용기인심편기술능쾌속감정분지간균균충，병구분MTC화NTM，구유간편쾌속급준학성、특이성、령민도고적우점。
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 .