KOD DNA聚合酶荧光PCR技术在肺结核诊断中的应用评价

Clinical evaluation of fluorescence probe PCR based on KOD DNA polymerase technology in the diagnosis of pulmonary tuberculosis

目的评估基于KOD DNA聚合酶的荧光PCR技术在肺结核诊断中的临床应用价值。方法采集2019年10月至2020年4月疑似肺结核的住院患者痰标本,进行涂片抗酸染色、MGIT960液体培养、KOD DNA聚合酶荧光PCR和Taq DNA聚合酶荧光PCR检测。以临床诊断结果为参照,MGIT960液体培养法为金标准,荧光PCR与Taq DNA聚合酶荧光PCR法比较,评价KOD DNA聚合酶荧光PCR法检测MTB的效能。结果共对337例临床样本进行检测,MGIT960液体培养法、Taq DNA聚合酶荧光PCR和KOD DNA聚合酶荧光PCR检测MTB的敏感度分别为71.56%(156/218)、76.15%(166/218)和76.61%(167/218)。KOD DNA聚合酶荧光PCR与MGIT960液体培养检测敏感度差异无统计学意义(χ2=1.445,P=0.229>0.05)。KOD DNA聚合酶荧光PCR检测MTB的敏感度和特异度分别为92.31%(144/156)和86.39%(146/169),涂阴患者中检测敏感度和特异度分别84.21%(64/76)和88.48%(146/165)。结论KOD DNA聚合酶荧光PCR检测MTB敏感度和特异度较高,能够为临床诊断肺结核提供依据,操作简便、快速,适合在我国各级结核病实验室推广。

To evaluate the clinical application value of fluorescence probe PCR based on KOD polymerase technology in the diagnosis of pulmonary tuberculosis(TB).Between October 2019 and April 2020,we collected a total of 337 samples from patients with suspected TB.Each specimen was examined with smears,MGIT960 solid culture,and fluorescence probe PCR based on KOD DNA polymerase and Taq DNA polymerase.Using the clinical diagnosis as a reference standard and MGIT960 solid culture as the gold standard,we compared the results with those of fluorescence probe PCR based on Taq DNA polymerase and evaluated the performance of fluorescence probe PCR based on KOD DNA polymerase.On the basis of the diagnosis,the sensitivities of MGIT960 solid culture,and fluorescence probe PCR based on Taq DNA polymerase and KOD DNA polymerase were 71.56%,76.15%and 76.61%,respectively.Fluorescence probe PCR based on KOD DNA polymerase produced similar results to those of MGIT960 solid culture(χ2=1.445,P=0.229,P>0.05).With the MGIT960 solid culture method result as the gold standard,the sensitivity and specificity of fluorescence probe PCR based on KOD DNA polymerase were 92.31%(144/156)and 86.39%(146/169),respectively;the sensitivity and specificity in smear negative patients were 84.21%(64/76)and 88.48%(146/165),respectively.Fluorescence probe PCR based on KOD DNA polymerase is easily performed and has relatively high sensitivity and specificity.This method can provide a basis for clinical diagnosis of pulmonary TB and is suitable for all types of laboratories.