恒温扩增荧光检测技术在基层结核病实验室的应用价值

Application of isothermal amplification fluorescence in peripheral tuberculosis laboratory

目的探讨在基层结核病实验室采用恒温扩增荧光检测技术检测可疑肺结核患者痰标本诊断结核病的价值。方法搜集河南省4个项目点县级结核病实验室2014-2015年期间可疑肺结核患者的痰标本4242例，分别进行痰涂片、痰培养和恒温扩增荧光检测技术检测，比较恒温扩增荧光检测技术和传统痰涂片、痰培养方法的敏感度、特异度等。计数资料采用χ2检验，以P〈0．05为差异有统计学意义。结果纳入的4242例患者中，痰涂片检出阳性者351例，阳性检出率为8．21％（351／4276）；痰培养检出阳性者576例，阳性检出率为13．47％（576／4276）；恒温扩增荧光检测技术检出阳性者733例，阳性检出率为17．14％（733／4276）。3种检测技术的阳性检出率比较，差异有统计学意义（χ2=153．412，P〈0．001）。以痰培养试验结果为标准，恒温扩增荧光检测技术的敏感度和特异度分别为86．81％（500／576）、93．64％（3433／3666）；阳性预测值为68．21％（500／733），阴性预测值为97．83％（3433／3509），与痰培养有较高的一致性（Kappa值为0．722）。痰涂片检测的敏感度为57．29％（330／576），特异度为99．43％（3645／3666），阳性预测值为94．02％（330／351），阴性预测值为93．68％（3645／3891）；Kappa值为0．679。痰涂片与恒温扩增荧光检测技术检测痰标本的敏感度比较，差异有统计学意义（χ2=153．336，P〈0．001）。结论恒温扩增荧光检测技术检测的敏感度和特异度均明显高于传统的痰涂片，且其具有检测快速、操作简便等优势，便于在我国基层结核病实验室中推广应用。

Objective To evaluate the isothermal amplification fluorescence technique in detecting Mycobacte- rium tuberculosis in sputum of suspects in peripheral tuberculosis laboratory. Methods Smear microscopy, L6wenstein-Jensen culture and isothermal amplification fluorescence tests were respectively performed to 4242 sputum samples from suspicious pulmonary tuberculosis patients in four county level laboratries in Henan province during 2014 to 2015, the sensitivity, specificity and detection time were compared with each method using Chi square test and the P value was set as 〈0. 05. Results Of 4242 TB suspects＇ samples, the number of smear positive cases 351 （8.21%）, the number of culture positive was 576 （13.47%）, the number of isothermal amplification fluores- cence 733 （17.14-/00）. The sensitivity of three detection methods were statistically significantly different,χ2 =153. 412,P〈0.001. The reporting time of the isothermal amplification fluorescence method was about 2 hours. Compared with culture as the standard, the sensitivity and specificity of the isothermal amplification fluorescence method were 86.81%（500/576） and 93.64% （3433/3666） respectively. Positive predictive value （PPV） and nega- tive predictive value （NPV） of the isothermal amplification fluorescence method were 68.21% （500/733） and 97.83 （3433/3509） respectively. The sensitivity, specificity, PPV and NPV of sputum smear microscopy were 57.29%（330/576）, 99.43% （3645/3666）, 94.02% （330/351）, 93. 68% （3645/3891） respectively. The results of the isothermal amplification fluorescence method and sputum smear microscopy was consistent with L6wenstein-Jensenculture （Kappa values were 0. 722 and 0. 679）. There was a statistically significantly difference in sensitivity between the isothermal amplification fluorescence method and sputum smear microscopy （χ2=153. 336, P〈0.01 ）. Conclusion The isothermal amplification fluorescence method shows more sensitivity and specificity than the tradi- tional methods, and is a rapid, sensitive, simple and specific method for the detection of Mycobacterium tuberculosis, then play an important role in diagnosing tuberculosis in peripheral laboratories.