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1 [ ]- - T- SPOT.TB T-SPOT.TB T-SPOT.TB86.5%(95%CI 71.2%~95.5%) 100%(95%CI 90.5%~100%) 52.9%(95%CI 27.8%~77.0%) 35.3%(95%CI 14.2%~61.7%) 80.0%(95%CI 64.4%~90.9%) 77.1%(95%CI 62.7%~88.0%) 64.3%(95%CI 35.1%~87.2%) 100%(95%CI 54.1%~100%)ROC T-SPOT.TB 47SFC/ % 88.2%- T-SPOT.TB T-SPOT.TB 47SFC/ [] - T-SPOT.TB [ ] R521.7 [ ] A [ ] (2018) [DOI] /j.issn tuberculous pleurisy ZHOU Min 1, YANG Qing-luan 2, CHEN Hua-xin 1, YU Zhi-ming 1, GAO Liang 1, LIU Qian-qian 2, OU Qin-fang 1* 1 Department of Pulmonary Diseases, Fifth People s Hospital of Wuxi, Wuxi, Jiangsu , China 2 Department of Infectious Diseases, Huashan Hospital affiliated to Fudan University, Shanghai , China * Corresponding author, @fudan.edu.cn This work was supported by the Project of Science and Technology of Wuxi (CSZ00N1229) [Abstract] Objective To evaluate the diagnostic value of interferon- release assay of blood and pleural effusion for tuberculous pleurisy. Methods Fifty-six adult patients with suspected tuberculous pleurisy were enrolled in our study. The blood and pleural effusion interferon- release assay were measured by T-SPOT.TB test in 38 pleural tuberculosis patients and 18 nontuberculous pleurisy controls. The diagnostic sensitivity, specificity, predictive value of T-SPOT.TB in pleural effusion mononuclear cells (PE-MC) and peripheral mononuclear cells (PBMC) were analyzed. Results The sensitivities and specificities, positive predictive values and negative predictive values, respectively, of the PE-MC and PBMC for diagnosing were as follows: 86.5%(95% confidence interval[ci] 71.2%-95.5%) and 100%(95%CI 90.5%-100%); 52.9%(95%CI 27.8%-77.0%) and 35.3%(95%CI 14.2%- 61.7%); 80.0%(95%CI 64.4%-90.9%) and 77.1%(95%CI 62.7%-88.0%); 64.3%(95%CI 35.1%-87.2%) and 100%(95%CI 54.1%- 100%). By ROC curve analysis, a cut-off value of 47SFC/ cells in PE-MC showed a sensitivity of 89.2% and a specificity of 88.2%. Conclusion T-SPOT.TB in PE-MC could be an accurate diagnostic method for tuberculous pleurisy in TB endemic settings. Moreover, 47SFC/ cells might be the optimal cut-off value for diagnosing tuberculous pleurisy. [Key words] interferon- release assay; T-SPOT.TB; pleural effusion; tuberculous pleurisy; diagnosis [ ] (CSZ00N1229) [ ] [ ] ( ) ( ) [ ] @fudan.edu.cn [1-2] [3-4] [5]

2 Med J Chin PLA, Vol. 43, No. 1, January 1, [6] [7] - (IFN- ) [8] T- SPOT.TB IFN- -6(early secretary antigenic target6 ESAT-6) -10(culture filtrate protein10 CFP-10) IFN- T [9] T-SPOT.TB ( ) () 1~3 < ml 50ml 1.2.2IFN- T-SPOT.TB(Oxford Immunotec Abingdon UK) (Mycobacterium tuberculosis)mtb 1(region of difference RD1) T 6h T-SPOT.TB ESAT-6(6 000kD) CFP-10(10 000kD) Ficoll-Hypaque (peripheral blood mononuclear cells PBMCs)(pleural effusion mononuclear cells PE-MCs) PBMCs PE-MCs - ( ) 37 5%CO 2 16~18h ELISPOT (spots forming cells SFC)6 2 PBMCs PE- MCs T-SPOT.TB 1.3SPSS 23.0 Mann-Whiney (Fisher's test) P< (30~72) (55~77) 15 (83.3%) 3 (16.7%) (P>0.05) (P=0.03) (P<0.01) ( 1) 2.2T-SPOT.TB T-SPOT.TB % 37 T- SPOT.TB 100% T- SPOT.TB % 17 T-SPOT.TB % T-SPOT.TB(P<0.05 1) T- SPOT.TB ESAT-6 CFP SFC/ T- SPOT.TB 86.5%(95%CI 71.2%~95.5%) 100%(95%CI 90.5%~100%) 52.9%(95%CI 27.8%~77.0%) 35.3%(95%CI 14.2%~61.7%) 80.0%(95%CI 64.4%~90.9%) 77.1%(95%CI 62.7%~88.0%) 64.3%(95%CI 35.1%~87.2%) 100%(95%CI

3 Tab.1 Comparison of various diagnostic index in 56 patients with suspected tuberculous pleurisy Item Tuberculous pleurisy (n=38) Non-tuberculous pleurisy (n=18) P value T-SPOT.TB [n(%)] PBMCs T-SPOT.TB 32/37(86.5) 8/17(47.1) PE-MCs T-SPOT.TB 37/37(100) 11/17(64.7) Blood [median (IQR)] WBC (10 9 /L) 5.57( ) 6.61( ) Neutrophil (%) 68.50( ) 71.25( ) Lymphocyte (%) 22.50( ) 19.00( ) Hb (g/l) ( ) ( ) PLT (10 9 /L) ( ) ( ) ESR (mm/h) 57.50( ) 38.00( ) CSF [median (IQR)] Protein (g/l) 40.95( ) 46.00( ) Lymphocyte (%) 84.00( ) 75.00( ) ADA (U/L) 42.60( ) 12.00( ) LDH (U/L) ( ) ( ) PBMCs. Peripheral blood mononuclear cells; PE-MCs. Pleural effusion mononuclear cells; WBC. White blood cell; Hb. Hemoglobin; PLT. Platelet; ESR. Erythrocyte sedimentation rate; ADA. Adenosine deaminase; LDH. Lactic dehydrogenase; IQR. Interquartile range IFN- producting T cell responses (SFC/ cells) IFN-ESAT-6 CFP-10 Fig.1 Box-and-whisker plot showing responses to ESAT-6 and CFP-10 according to the PBMC and PE-MC T-SPOY.TB PBMCs. Peripheral blood mononuclear cells; PE-MCs. Pleural effusion mononuclear cells; TB. Tuberculous pleurisy; non-tb. Nontuberculous pleurisy; A. ESAT-6; B. CFP-10; P<0.05 A B A B A B A B TB non-tb TB non-tb PBMCs PE-MCs 54.1%~100% 2) T- SPOT.TB 2.3 ROCT- SPOT.TB ROC 58 T- SPOT.TB T- SPOT.TB ROC (AUC) (95%CI 0.891~1.000 P<0.001) T-SPOT.TB AUC( %CI 0.666~0.860 P < )ROC T- SPOT.TB 47SFC/ % 88.2% 94.3% 78.9% T-SPOT.TB 97.4% 90.9% T-SPOT.TB (%) Tab.2 Diagnostic performance of PBMC and PE-MC T-SPOT.TB in the study population (%) T-SPOT.TB Sensitivity Specificity PPV NPV LR + LR PBMCs>6SFC ( ) 52.9( ) 80.0( ) 64.3( ) 1.84( ) 0.255( ) PE-MCs>6SFC 100.0( ) 35.3( ) 77.1( ) 100.0( ) 1.55( ) PE-MCs>47SFC ( ) 88.2( ) 94.3( ) 78.9( ) 7.58( ) 0.123( ) PBMCs>6SFC or PE-MCs>6SFC 100.0( ) 29.4( ) 75.5( ) 100.0( ) 1.42( ) PBMCs>6SFC or PE-MCs>47SFC 97.4( ) 55.6( ) 82.2( ) 90.9( ) 2.19( ) 0.047( ) PBMCs>6SFC and PE-MCs>6SFC 86.1( ) 56.3( ) 81.6( ) 64.3( ) 1.97( ) 0.247( ) PBMCs>6SFC and PE-MCs>47SFC 73.7( ) 88.9( ) 93.3( ) 61.5( ) 6.63( ) 0.296( ) PBMCs. Peripheral blood mononuclear cells; PE-MCs. Pleural effusion mononuclear cells; SFC. Spots forming cells; PPV. Positive predictive value; NPV. Negative predictive value; LR +. Positive likelihood ratio; LR. Negative likelihood ratio. 1 SFC is short for SFCs / mononuclear cells; 2 Cut-off derived from the area under the curve by the Youden index

4 Med J Chin PLA, Vol. 43, No. 1, January 1, Sensitivity specificity 2T-SPOT.TBROC Fig.2 ROC curves in PBMCs and PE-MCs T-SPOT.TB for diagnosis of tuberculous pleurisy ( 2) PBMC (0.7663) PE-MC (0.9459) T-SPOT.TB T-SPOT.TB T-SPOT.TB 6SFC/ IFN- MtbT T T Souza [10] T Barnes [11] CD4 + T T IFN- IFN- T-SPOT.TBT-SPOT.TB IFN- 93% [12] 6SFC/ T-SPOT.TB T-SPOT.TB 47SFC/ %89.2% 52.9%88.2% 80.0%94.3% 64.3%78.9% T- SPOT.TB 6SFC/ T- SPOT.TB [9] 187 T- SPOT.TB T- SPOT.TB 6SFC/ T-SPOT.TB ROC T- SPOT.TB AUC T- SPOT. TB(0.7663) T-SPOT.TB 47SFC/ T-SPOT.TB 89.2% 88.2% Zhang [13] 98 T-SPOT.TB (94.5%) (76.1%) 54 SFC/ T-SPOT.TB IFN- T-SPOT.TB T-SPOT.TB 47SFC/ T-SPOT.TB [1] Hooper CE, Lee YC, Maskell NA. Interferon-gamma release assays for the diagnosis of TB pleural effusions: hype or real hope?[ J]. Curr Opin Pulm Med, 2009, 15(4): [2] Shi XP, Wang J, Wang X, et al. Clinical application of T-spot test of Mycobacterium tuberculosis infection for diagnosis of suspected pulmonary tuberculosis patients[ J]. Med J Chin PLA, 2017, 42(11): [,,,. T [ J]., 2017, 42(11): ] [3] Porcel JM. Tuberculous pleural effusion[ J]. Lung, 2009, 187(5): [4] Liu HM, Chu Z, Tian R, et al. Application of IL-10 and IL-12 detection in differential diagnosis of tuberculous and malignant pleural effusion[ J]. J Jilin Univ (Med Ed), 2017, 43(4): , 861. [,,,. IL-10 IL-12 [ J]. ( ), 2017, 43(4): , 861.] [5] Greco S, Girardi E, Masciangelo R, et al. Adenosine deaminase and interferon gamma measurement for the diagnosis of tuberculous pleurisy: a meta-analysis[ J]. Int J Tuberc Lung Dis,

5 , 7(8): [6] Ou QF, Gao Y, Shao LY, et al. Assessment the diagnostic power by interferon-gamma release of the whole blood and pleural biopsy in tuberculous pleurisy[ J]. Chin J Exp Clin Infect Dis, 2014, 8(3): [,,,. [J]., 2014, 8(3): ] [7] Pai M, Zwerling A, Menzies D, et al. Systematic review: T-cellbased assays for the diagnosis of latent tuberculosis infection: an update[ J]. Ann Intern Med, 2008, 149(3): [8] Liu Y, Ou M, He S, et al. Evaluation of a domestic interferongamma release assay for detecting Mycobacterium tuberculosis infection in China[ J]. Tuberculosis (Edinb), 2015, 95(4): [9] Liu XQ. The clinical application of IGRA test in body fluid (serositis and cerebral spinal fluid) for diagnosis of tuberculosis[ J]. Chin J Antituberc, 2015, 37(7): [. -() [ J]., 2015, 37(7): ] [10] Souza MC, Penido C, Costa MF, et al. Mechanisms of T-lymphocyte accumulation during experimental pleural infection induced by Mycobacterium bovis BCG[ J]. Infect Immun, 2008, 76(12): [11] Barnes PF, Mistry SD, Cooper CL, et al. Compartmentalization of a CD4 + T lymphocyte subpopulation in tuberculous pleuritis[ J]. J Immunol, 1989, 142(4): [12] L i u Y, O u, Q, Z h e n g, J, et al. A combination of the QuantiFERON-TB gold in-tube assay and the detection of adenosine deaminase improves the diagnosis of tuberculous pleural effusion[ J]. Emerg Microbes Infect, 2016, 5(8): e83. [13] Zhang L, Zhang Y, Shi X, et al. Utility of T-cell interferon-gamma release assays for diagnosing tuberculous serositis: a prospective study in Beijing, China[ J]. PLoS One, 2014, 9: e ( ) ( )

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