CAJ | 학술논문

目的：探讨针对hspX 基因设计引物的环介导等温扩增技术( LAMP)对结核性胸膜炎患者结核分枝杆菌的诊断价值。方法采用LAMP技术对结核分枝杆菌标准株 H37Rv的DNA 样品进行检测,评价LAMP技术的检测限；同时对8种非结核分枝杆菌菌株及阴沟肠杆菌和大肠埃希菌标准菌株进行鉴定,评价该方法的特异性。对58例结核性胸膜炎患者和32例肺癌患者胸腔积液样本结核分枝杆菌进行检测,评价LAMP 技术的临床应用价值。结果 LAMP对倍比稀释的H37Rv的DNA 样品检测结果为阳性,检测限为320aM。八种非结核分枝杆菌和2种普通菌菌株检测结果为阴性。 LAMP对胸腔积液标本检测的灵敏度是75．8%(44/58),定量PCR 为79．3%(46/58),无显著差别(χ2=0．25,P>0．05)；均显著高于涂片法39．6%(23/58),差异具有统计学意义(χ2=17．39,P<0．01)。结论 LAMP技术是一种简单快速有效的等温扩增技术,由于其较高的灵敏度和特异度,可以对结核性胸膜炎患者胸腔积液标本快速鉴定。
목적：탐토침대hspX 기인설계인물적배개도등온확증기술( LAMP)대결핵성흉막염환자결핵분지간균적진단개치。방법채용LAMP기술대결핵분지간균표준주 H37Rv적DNA 양품진행검측,평개LAMP기술적검측한；동시대8충비결핵분지간균균주급음구장간균화대장애희균표준균주진행감정,평개해방법적특이성。대58례결핵성흉막염환자화32례폐암환자흉강적액양본결핵분지간균진행검측,평개LAMP 기술적림상응용개치。결과 LAMP대배비희석적H37Rv적DNA 양품검측결과위양성,검측한위320aM。팔충비결핵분지간균화2충보통균균주검측결과위음성。 LAMP대흉강적액표본검측적령민도시75．8%(44/58),정량PCR 위79．3%(46/58),무현저차별(χ2=0．25,P>0．05)；균현저고우도편법39．6%(23/58),차이구유통계학의의(χ2=17．39,P<0．01)。결론 LAMP기술시일충간단쾌속유효적등온확증기술,유우기교고적령민도화특이도,가이대결핵성흉막염환자흉강적액표본쾌속감정。
Objective To explore the application of loop-mediated isothermal amplification ( LAMP) assay in the rapid diagnosis of patients with tuberculous pleuritis. Methods Mycobacterium tuberculosis in pleural effu-sion from 58 patients with tuberculous pleuritis and 32 lung cancer patients were tested by LAMP. The specificity of this assay was evaluated using H37Rv reference strains of mycobacterium tuberculosis, 8 species of non-tuberculous mycobacteria (NTM), eterobacter cloacae, and E. coli. Results H37Rv reference strain was successfully detected by this method, and there were no false-positive results of NTM, eterobacter cloacae and E. coli. The sensitivity of LAMP in detection of purified DNA from H37Rv was 320 aM. The detection sensitivity for the pleural effusion from 58 tuberculous pleuritis was 75. 8% (44/58), which was higher than smear microscopy (39. 6%). There was no significant difference between the sensitivity of LAMP and that of quantitative real-time PCR (χ2 =0. 25, P>0. 05). Conclusion Because of its speed, simplicity, sensitivity and specificity, the TB hspX LAMP assay is a potential gene diagnostic method for mycobacterium tuberculosis in patients with tuberculous pleuritis.