CAJ | 학술논문

目的：建立实时荧光环介导等温扩增快速检测结核分枝杆菌的方法。方法针对结核分枝杆菌特异序列建立实时荧光环介导等温扩增技术；对其特异性和敏感性进行评估；通过敏感性和特异性等指标验证其稳定性。结果建立的实时荧光环介导等温扩增法对结核和非结核分枝杆菌标准菌株检测的敏感性和特异性均为100%，灵敏度可达102个菌/mL；痰标本结核分歧杆菌检测的敏感性为94.64%（53/56），荧光定量聚合酶链式反应方法检测的敏感性为92.86%(52/56)；检测的特异性为96.55%（28/29），荧光定量聚合酶链式反应方法检测的特异性为96.55%(28/29)。结论建立实时荧光环介导等温扩增检测结核分枝杆菌的方法，敏感性和特异性强，灵敏度高，稳定性好，可用于痰液中结核分枝杆菌的快速检测。
목적：건립실시형광배개도등온확증쾌속검측결핵분지간균적방법。방법침대결핵분지간균특이서렬건립실시형광배개도등온확증기술；대기특이성화민감성진행평고；통과민감성화특이성등지표험증기은정성。결과건립적실시형광배개도등온확증법대결핵화비결핵분지간균표준균주검측적민감성화특이성균위100%，령민도가체102개균/mL；담표본결핵분기간균검측적민감성위94.64%（53/56），형광정량취합매련식반응방법검측적민감성위92.86%(52/56)；검측적특이성위96.55%（28/29），형광정량취합매련식반응방법검측적특이성위96.55%(28/29)。결론건립실시형광배개도등온확증검측결핵분지간균적방법，민감성화특이성강，령민도고，은정성호，가용우담액중결핵분지간균적쾌속검측。
Objective To establish a rapid method of real-time fluorescence loop-mediated isothermal amplification (LAMP) for detection of Mycobacterium tuberculosis. Methods According to the specific sequence of IS6110 in Mycobacterium tuberculosis, 6 primers of loop-mediated isothermal amplification were designed and synthesized. With the ESE-Quant tube scanner platform, a real-time fluorescence loop-mediated isothermal amplification was established. 14 strains of non-tuberculous mycobacteria standard strains and 14 strains of Mycobacterium tuberculosis isolated from cases were used to evaluate specificity and sensitivity of the method. Sputum samples were collected from 56 positive cases for Mycobacterium tuberculosis and 29 negative cases for Mycobacterium tuberculosis. The stability of the method was validated by the sensitivity and specificity compared to the real-time fluorescent PCR method. Results Real-time fluorescence loop-mediated isothermal amplification method was established, the specificity and sensitivity of Mycobacterium tuberculosis and non-tuberculous mycobacterial standard strains reflected 100% . In 56 cases of sputum with Mycobacterium tuberculosis, the sensitivity of real-time fluorescence LAMP method was 94.64% (53/56), while the sensitivity of real-time fluorescence polymerase chain reaction (PCR) method was 92.86% (52/56). In 56 cases of sputum without Mycobacterium tuberculosis, the specificity of real-time fluorescence LAMP method was 96.55% (28/29), while the specificity of real-time fluorescence PCR method was 96.55% (28/29). Conclusion The method of real-time fluorescence loop-mediated isothermal amplification (LAMP) for detection of Mycobacterium tuberculosis with ESE-Quant tube scanner platform was established, which has strong specificity, high sensitivity, good stability and can be used for rapid detection of Mycobacterium tuberculosis in sputum.