环介导等温扩增技术在肺结核病诊断中的应用评价

Evaluation of loop-mediated isothermal amplification assay in the diagnosis of pulmonary tuberculosis

目的评估环介导等温扩增技术(loop-mediated isothermal amplification,LAMP)在肺结核病诊断中的应用价值。方法选取2016年1月-2018年3月陕西省结核病防治研究所和汉中市第二人民医院就诊的258例肺结核可疑症状者作为研究对象,同时应用痰涂片镜检法、固体培养法、LAMP对所有研究对象的同一份痰样本进行检测。计算3种方法的阳性检出率。以临床诊断结果为标准法,比较痰涂片镜检法、固体培养法和LAMP检测结核分枝杆菌(mycobacterium tuberculosis,MTB)的效能;在涂阴患者中,比较固体培养法和LAMP检测MTB的效能;涂阴培阴患者中,评估LAMP检测MTB的效能。结果在唾液痰标本中,LAMP的阳性检出率34.61%显著高于痰涂片镜检法12.82%和固体培养法19.23%的阳性检出率,差异均有统计学意义(χ2=10.24,P=0.001;χ2=4.69,P=0.03)。以临床诊断为标准,LAMP的敏感度73.30%显著高于痰涂片镜检法46.02%和固体培养法58.52%的敏感度,差异均有统计学意义(χ2=25.98,P<0.001;χ2=8.55,P=0.003),LAMP、痰涂片镜检法、固体培养法的阳性检出率分别为51.55%、31.40%、39.92%,Kappa值分别为0.60、0.35、0.47;在涂阴患者中,LAMP的敏感度52.63%,显著高于固体培养法的敏感度33.68%,差异有统计学意义(χ2=6.95,P=0.01),LAMP、固体培养法的阳性检出率分别为30.51%、18.08%,Kappa值分别为0.32、0.31;在涂阴培阴患者中,LAMP的敏感度和特异度分别为39.68%、95.12%,Kappa值为0.37。结论 LAMP对唾液痰标本的阳性检出率高于传统检测方法。LAMP具有较高的敏感度和特异度,对肺结核病尤其是涂阴肺结核病的快速诊断具有一定的诊断价值。

Objective To evaluate diagnostic value of loop-mediated isothermal amplification (LAMP)technology in the diagnosis of pulmonary tuberculosis.Methods A total of 258 pulmonary tuberculosis suspects were continuously recruited in Shaanxi provincial Institute for Tuberculosis Control and Prevention and The second people's Hospital of Hanzhong from January 2016 to March 2018,the same specimens were detected by smear,solid culture and LAMP at the same time.The positive detection rate of the 3 methods was calculated.Based on the clinical diagnosis result,com- pare efficiency of smear,solid culture and LAMP in all patents;compare efficiency of solid culture and LAMP in smear negative patients and evaluate efficiency of the LAMP in both of smear and solid culture negative patients.Results The postive detection rate of LAMP (34.610%)was significantly higher than that of smear (12.82%) and solid culture (12.820%),the difference being significant(X^2=10.24,P =0.001;X^2=4.69,P=0.03).Using clinical diagnosis result as the gold standard,the sensitivities of LAMP(73.30%)was significantly higher than that of smear(46.02%)and solid culture(58.52%),the difference being significant(X^2=25.98,P<0.001;X^2 =8.55,P=0.003).The positive de- tection rates of LAMP,smear and solid culture were 51.55%,31.40%,39.92% respectively.Kappa value were 0.60, 0.35 and 0.47,respectively;the sensitivities of LAMP(52.63%)was significantly higher than that of solid culture (33.68%)in smear negative patients,the difference being significant(X^2=6.95,P=0.01).The positive detection rates of LAMP and solid culture Were 30.51% and 18.08%,respectively.Kappa value were 0.46 and 0.32,respectively;the sensitivity and specificity of LAMP were 39.68% and 95.12% in both of smear and solid culture negative patients,re- spectively.The positive detection rates of LAMP was 20.00%,Kappa value was 0.37.Conclusion The postive detec- tion rate of LAMP was higher than that of bacteriological testing.LAMP has a certain diagnostic value for rapid diagno-sis of pulmonary tuberculosis with high sensitivity and specificity,especially for smear negative.