中国人兽共患病学报2025，Vol.41Issue(10)：1034-1039,6.DOI:10.3969/j.issn.1002-2694.2025.00.167
荧光PCR法对结核分枝杆菌复合群和利福平耐药基因检测性能评价
Performance of fluorescence PCR in detecting Mycobacterium tuberculosis complex and rifampicin resistance
摘要
Abstract
The diagnostic value of Mycobacterium tuberculosis(MTB)complex and rifampicin resistance test kits(fluorescence PCR method)in detecting for MTB complex and rifampicin resistancein sputum samples was evaluated.A total of 271 patients with suspected tuberculosis were prospectively and consecutively enrolled at Hunan Chest Hospital between April 1,2024,and November 30,2024.Of these,229 patients were confirmed to have tuberculosis,whereas 42 patients were not-tuberculosis samples were col-lected from all patients and subjected to fluorescence PCR,Xpert MTB/RIF(abbreviated as Xpert),and MGIT 960 culture and drug sensitivity testing.Clinical diagnosis and MTB culture results served as reference standards for TB diagnosis,whereas phenotypic drug susceptibility testing and Xpert served as reference standards to for assessment of rifampicin resistance.The sensitivity,specificity,positive predictive value,and negative predictive value of the fluorescence PCR method were analyzed.Kappa tests were performed to analyze the concordance between detection techniques.With clinical diagnosis as the gold standard,the sensitivity and specificity of the fluorescence PCR method for the diagnosis of TB were 65.1%(149/229)and 97.6%(41/42),and the consistency test for the fluo-rescence PCR and Xpert methods showed high consistency(Kappa value=0.993).With the MGIT 960 liquid culture as the reference standard,the positive detection rate of the fluorescence PCR method for the detection of patients with positive cultures was 91.9%(102/111,95%CI:85.2%-96.2%),and the positive detection rate for 147 patients with sputum culture-negative TB was 27.9%(41/147,95%CI:21.3%-35.6%).With the phenotypic drug sensitivity test as the gold standard,the sensitivity and specificity of the fluo-rescence PCR method for the detection of rifampicin resistance were 100.0%(31/31)and 96.6%(28/29)respectively,and the consis-tency between tests was high(Kappa value=0.967).With Xpert as the gold standard,the sensitivity and specificity of fluorescence PCR for the detection of rifampicin resistance were 95.8%(46/48)and 99.0%(99/100),respectively,and the consistency between tests was high(Kappa value=0.953).Finally,samples with rifampicin resistance detected with the fluorescence PCR method had a clear rpoB gene mutation type according to one-generation sequencing.In conclusion,the fluorescence PCR method showed high sen-sitivity in detecting MTB complex groups and rifampicin resistance,and had high concordance with Xpert.Therefore,this technique can serve a rapid test for TB diagnosis to increase the rate of positive TB pathology and detection of rifampicin resistance.This method is particularly suitable for use in lower-income countries and economically disadvantaged grassroots communities.关键词
肺结核/痰液/诊断/结核分枝杆菌复合群/利福平耐药Key words
pulmonary tuberculosis/sputum/diagnosis/Mycobacterium tuberculosis complex/rifampicin resistance分类
医药卫生引用本文复制引用
刘彬彬,万小洁,谭歆昀,王珏,郭婧玮,李文彬,钟彪,谭云洪..荧光PCR法对结核分枝杆菌复合群和利福平耐药基因检测性能评价[J].中国人兽共患病学报,2025,41(10):1034-1039,6.基金项目
湖南省卫生健康高层次人才重大专项(No.R2023098) （No.R2023098）
Hunan Provincial Major Special Project for High-Level Talents in Health(No.R2023098) （No.R2023098）
