Value of bronchoalveolar lavage fluid metagenomic next-generation sequencing to the diagnosis of pulmonary tuberculosis
Objective To compare the efficiencies of bronchoalveolar lavage fluid(BALF)metagenomic next-generation sequencing(mNGS),Xpert and acid-fast staining on the diagnosis of pulmonary tuberculosis,and to explore the diagnostic value of BALF mNGS for pulmonary tuberculosis.Methods Totally 265 patients with suspected pulmonary tuberculosis received bronchoscopy,and BALF mNGS,Xpert and acid-fast staining in Henan Provincial People's Hospital from May 2021 to July 2023,and were divided into pulmonary tuberculosis group(n=34)and non-pulmonary tuberculosis group(n=231)according to the etiology and histopathological results.The clinical data as age and pulmonary comorbidities as well as the mixed infectious pathogenic microorganisms detected by mNGS were compared between two groups.The detection rates of Mycobacterium tuberculosis complex by BALF mNGS,Xpert and acid-fast staining,as well as their sensitivities and specificities in the diagnosis of pulmonary tuberculosis were calculated.The low sequence number detected by BALF mNGS was recorded in pulmonary tuberculosis group,and the numbers of mNGS sequences were compared between patients with positive and negative Xpert and acid-fast staining.Results The patients were younger in pulmonary tuberculosis group[47.5(30.0,70.5)years]than in non-pulmonary tuberculosis group[59.0(50.0,69.0)years](Z=2.317,P=0.021),and there were no significant differences in the pulmonary comorbidities,extrapulmonary comorbidities,and mixed infectious pathogenic microorganisms detected by BALF mNGS between two group(P＞0.05).In 34 patients with pulmonary tuberculosis,there were 29 cases of BALF mNGS positive,24 of Xpert positive,11 of acid-fast staining positive,8 cases of mNGS+Xpert+acid-fast staining positive,22 of both mNGS and Xpert positive,and 8 of both mNGS and acid-fast staining positive.The detection rate of Mycobacterium tuberculosis complex was higher by BALF mNGS(10.9％)and Xpert(9.1％)than that by acid-fast staining(4.2％)(x2=8.654,P=0.003;x2=6.050,P=0.012),and showed no significant difference between BALF mNGS and Xpert(x2=0.900,P=0.343).The sensitivity of BALF mNGS(85.3％)and Xpert(70.6％)was higher than that of acid-fast staining(32.4％)in diagnosing pulmonary tuberculosis(x2=12.042,P＜0.001;x2=8.471,P=0.002),and there was no significant difference between BALF mNGS and Xpert(x2=1.778,P=0.180).There were no significant differences in the specificities of BALF mNGS,Xpert and acid-fast staining in diagnosing pulmonary tuberculosis(100.0％,99.6％,99.1％)(x2=2.000,P=0.368).The sensitivity of BALF mNGS combined with Xpert in diagnosing pulmonary tuberculosis(91.2％)was higher than that of Xpert(x2=5.143,P=0.016),and showed no significant difference compared with BALF mNGS(x2=0.500,P=0.500).The specificity of BALF mNGS combined with Xpert in diagnosing pulmonary tuberculosis(99.6％)had no significant differences compared with BALF mNGS and Xpert(x2＜0.001,P＞0.999;x2＜0.001,P＞0.999).In pulmonary tuberculosis group,there were 4 cases with low mNGS sequence number(0 to 3),of which 2 were positive for Xpert,and acid-fast staining was all negative.The mNGS sequence number was higher in Xpert-positive patients[798.0(8.8,83 139.0)]than that in Xpert-negative patients[9.5(0,205.3)](Z=2.423,P=0.015),and showed no significant difference between the positive acid-fast staining patients[66 137.0(0,361 759.8)]and the negative acid-fast staining patients[22.5(3.3,747.3)](Z=1.515,P=0.130).Conclusion BALF mNGS technology has higher efficiency on diagnosing pulmonary tuberculosis than acid-fast staining,and BALF mNGS combined with Xpert can improve the diagnostic sensitivity of pulmonary tuberculosis.
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