环介导核酸等温扩增技术对疑似结核病患者的筛查效果

Screening effect of loop-mediated isothermal amplification for suspected tuberculosis

目的评价环介导核酸等温扩增（100p-mediated isothermal amplification，LAMP）技术对疑似结核病患者的筛查效果。方法选取沈阳市第十人民医院2015年5月至2016年6月收治的疑似结核病患者604例，收集标本604份，其中痰标本503份，胸腔积液抽取液98份，支气管肺泡灌洗液3份，分别行涂片抗酸染色镜检（简称“涂片镜检”）、改良罗氏固体培养（简称“固体培养”）、MGIT960液体培养（简称“液体培养”）和LAMP检测，以临床诊断结果为标准，评价4种检测技术对结核病患者的检测效能，采用x^2检验比较结核分枝杆菌的阳性检出率，以P（0．05为差异有统计学意义。结果以临床诊断结果为参照，LAMP检测、涂片镜检、液体培养、固体培养的敏感度分别为72．78％（262／360）、51．67％（186／360）、66．67％（240／360）和59．17％（213／360）；特异度分别为88．11％（215／244）、99．59％（243／244）、99．59％（243／244）和99．59％（243／244），并且具有良好的一致性（Kappa值依次为0．58、0．46、0．61和0．54）。LAMP检测对病程〈1年患者的阳性检出率[45．76％（189／413）]明显高于涂片镜检E27．12％（112／413）1、液体培养[-37．05％（153／413）]和固体培养[-31．48％（130／413）（x^2=62．41、16．62、40．95，P值均=0．000）。当患者出现的临床症状45个时，LAMP的阳性检出率（44．37％，268／604）明显高于涂片镜检（28．15％，170／604）、液体培养法（36．75％，222／604）和固体培养法（32．12％，194／604）（x^值分别为76．22、21．59和47．21，P值均=0．000）。LAMP检测对涂片镜检阴性，液体培养及固体培养阴性标本具有较高的阳性检出率[分别为28．78％（120／417），（21．49％（78／363）和25．90％（101／390）]。结论LAMP技术检测结核分枝杆菌较涂片镜检、液体培养和固体培养检测效能高，适用于病程短、临床症状较轻（1～5个症状）、涂阴和培阴的疑似结核病患者的筛查。

Objective To evaluate screening effect of loop-mediated isothermal amplification assay （LAMP） for suspected tuberculosis. Methods A total of 604 suspected tuberculosis patients from Shenyang 10th People＇s Hospital between May 2015 and June 2016 were selected, and 604 specimens were collected, including 503 sputum, 98 pleural effusions and 3 bronchoalveolar lavage fluid. All specimens were examined by smear microscopy, solid culture, MGIT 960 liquid culture and LAMP, in order to test the different efficacy of four methods, based on clinical diagnosis. The positive detection rate was analyzed by x^2 test, and P〈0.05 was considered statistically signifi- cant. Results Compared with clinical diagnosis, sensitivities of these four methods were 72.78% （262/360）, 51.67% （186/360）, 66.67%（240/360） and 59.17% （213/360）, respectivelyand the specificities were 88.11% （215/244）, 99.59% （243/244）, 99.59% （243/244） and 99.59% （243/244）, what＇s more, the consistency was good （Kappa value were 0. 58, 0. 46, 0. 61 and 0. 54, respectively）. The positive detection rate of LAMP in patients with short course （less than 1 year） （45.76% （189/413）） was significantly higher than those of smear microscopy （27.12% （112/413））, MGIT 960 culture （37.05% （153/413）） and solid culture （31.48% （130/413）） （x^2=62. 41, 16.62, 40.95, respectively, all P=0. 000）. In patients with less than 5 symptoms, positive detection rate of LAMP （44. 37% （268/604）） was significantly higher than those of microscopy （28. 15% （170/604））, liquid culture （36.75% （222/604）） and solid culture （32.12% （194/604）） （x^2=76.22, 21.59 and 47.21, respectively,all P=0. 000）. For smear negative, liquid culture negative and solid culture negative specimens, LAMP also had high positive detection rate （28.78% （120/417）, 21.49% （78/363） and 25. 90% （101/390）, respectively）. Conclusion LAMP is more effective than microscopy, liquid culture and solid culture. It is suitable for the screening of suspected tuberculosis patients with short course, mild clinical symptoms （less than 5 symptoms）, smear negative and culture negative.