联合应用Xpert MTB/RIF、SAT-TB及MTB/NTM PCR方法检测支气管肺泡灌洗液对痰涂阴性肺结核的快速诊断价值

Value of Xpert MTB/RIF,RNA isothermal amplification real‐time testing and MTB/NTM PCR combined with detection of bronchoalveolar lavage fluid in rapid diagnosis of sputum smear‐negative pulmonary tuberculosis

目的评估联合应用多色巢式荧光定量PCR(Xpert mycobacterium tuberculosis and rifampicin sensitivity test,Xpert MTB/RIF)、RNA恒温扩增实时荧光检测(simultaneous amplification and testing of tuberculosis,SAT‐TB)、荧光探针分枝杆菌核酸检测(mycobacterium tuberculosis/non-mycobacterium tuberculosis polymerase chain reaction,MTB/NTM PCR)3种方法检测疑诊肺结核患者的支气管肺泡灌洗液标本对痰涂阴性肺结核的快速诊断价值。方法采集2019年10月至2021年9月杭州市红十字会医院收治的痰涂片至少3次阴性的疑诊肺结核的898例患者支气管肺泡灌洗液,以MIGT960液体培养阳性为金标准,评估Xpert MTB/RIF、SAT-TB及TB/NTM PCR在检出MTB感染、耐药结核感染以及NTM感染中的快速诊断价值。结果在对MTB的检出效能方面,Xpert MTB/RIF、SAT-TB及TB/NTM PCR对MTB的检出率分别为87.24%、80.73%、88.02%,3者均达到80%以上,其中SAT-TB的检出率最低,其余两者相当,差异有统计学意义(P<0.001)。而3者组合的平行试验对MTB的检出率最高达97.14%,差异有统计学意义(P<0.001)。在对耐药结核的检出效能方面,MIGT960液体培养及Xpert MTB/RIF分别检出8.59%及10.94%的耐药结核,虽Xpert较高,但差异无统计学意义(P=0.122);而两者组合的平行试验可以检出12.24%的耐药结核,显著高于单用Xpert的效能(P=0.019)。在对NTM的检出方面,MIGT960液体培养(14.03%)高于NTM-DNA(8.24%),差异有统计学意义(P<0.001),两者组合的平行试验的检出率更高(14.48%)。结论与传统细菌学检测方法相比,联合使用Xpert MTB/RIF、SAT-TB及MTB/NTM PCR可以快速、敏感地检测出痰涂阴性疑诊肺结核患者支气管肺泡灌洗液标本中的结核核酸,同步检出耐药,还同时可以鉴别MTB与NTM,缩短了确诊时间,给疑诊肺结核患者的早期快速诊断提供了有效的实验室方案,值得临床推广应用。

Objective To evaluate the value in detection of bronchoalveolar lavage fluid with Xpert mycobacterium tuberculosis and rifampicin sensitivity test(Xpert MTB/RIF),RNA simultaneous amplification and testing of tuberculosis(SAT-TB)and mycobacterium tuberculosis/non-mycobacterium tuberculosis polymerase chain reaction(MTB/NTM PCR)combined in diagnosis of sputum smear‐negative pulmonary tuberculosis.Methods From October 2019 to September 2021,the bronchoalveolar lavage fluid specimens were collected from patients with suspected pulmonary tuberculosis who were tested negative for sputum smear for at least 3 times,With Bactec MGIT 960 culture set as the golden standard,rapid diagnostic value of Xpert MTB/RIF combined with SAT‐TB and MTB/NTM PCR in testing MTB infection,drug-resistant TB infection,and NTM infection were evaluated and compared with single Xpert MTB/RIF,SAT-TB and MTB/NTM PCR.Results A total of 898 cases were positive cultured with Bactec MGIT 960 system,130 specimens were NTM,others were MTB.In the detection of tuberculosis,Xpert MTB/RIF,SAT-TB and MTB/NTM PCR could detect 670 specimens,620 specimens and 676 specimens respectively,detection rate were 87.24%,80.73%and 88.02%respectively;there was no significant difference in the value of diagnosis between the single detection of Xpert MTB/RIF and MTB/NTM PCR;the detection rate of SAT-TB was significantly lower than other two(P<0.001),detection rate of 3 combined parallel test(single positive)was significantly higher than Xpert or MTB/NTM PCR(P<0.001).Conclusion As compared with traditional bacteriological detection,using Xpert MTB/RIF,SAT-TB and MTB/NTM PCR in combination is rapid and sensitive in diagnosis of sputum‐negative drug-sensitive and-resistance pulmonary tuberculosis and can discriminate from MTB to NTM,shorten the diagnosis time,raise the accuracy,and offer an effective auxiliary mean for diagnosis of the sputum smear‐negative pulmonary tuberculosis,and it is worthy to be promoted in hospital.