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目的探讨环介导等温扩增法(loop-mediated isothermal amplification,LAMP)快速检测结核分枝杆菌技术在基层结防机构推广应用的可行性。方法选取深圳市宝安区疑似肺结核初诊患者300例,分别采用抗酸染色涂片法、L-J罗氏培养法及LAMP法检测痰标本中结核分枝杆菌。采用卡方检验和Kappa检验分析三种检测方法的一致性,以P<0.05为差异有统计学意义。结果 300例痰标本中,抗酸染色涂片法、L-J改良罗氏培养法和LAMP法的阳性检出率分别为13.7%(41/300)、20.7%(62/300)和22.0%(66/300);LAMP法灵敏度、特异度和诊符率分别为44.2%(61/138)、96.9%(157/162)、72.3%(218/300)。同改良罗氏培养法相比,Kappa值=0.94,P=0.219,差异无统计学意义。与涂片法相比,Kappa值=0.674,P<0.001差异有统计学意义。结论 LAMP法用于痰标本中结核分枝杆菌检测,其效能同罗氏培养法相当,同直接涂片法相比,LAMP法能够大幅提高阳性检出率,且技术操作较简便,具有较好的临床应用和推广前景。
Objective To investigate the feasibility of rapid detection of Mycobacterium tuberculosis by loop-mediated isothermal amplification (LAMP) in the popularization and application of grass-roots units. Methods 300 patients with newly diagnosed pulmonary tuberculosis in Bao’an District of Shenzhen City were enrolled in this study. Mycobacterium tuberculosis was detected by acid-fast staining, L-J Roche culture and LAMP respectively. Using chi-square test and Kappa test analysis of the consistency of the three detection methods, P <0.05 was considered statistically significant. Results The positive detection rates of acid-fast staining, LJ-modified Roche and LAMP in 300 sputum samples were 13.7% (41/300), 20.7% (62/300) and 22.0% (66 / 300). The sensitivity, specificity and consultation rate of LAMP were 44.2% (61/138), 96.9% (157/162) and 72.3% (218/300), respectively. Compared with the modified Roche culture method, Kappa value = 0.94, P = 0.219, the difference was not statistically significant. Compared with the smear method, Kappa value = 0.674, P <0.001 difference was statistically significant. Conclusions LAMP method is used to detect Mycobacterium tuberculosis in sputum samples. Its efficiency is comparable to that of Roche culture method. Compared with direct smear method, LAMP method can greatly improve the positive detection rate, and the technique operation is relatively simple and has a good clinical Application and promotion of the future.