Evaluation of real-time PCR of patient pleural effusion for diagnosis of tuberculosis
Tóm tắt
Pleural tuberculosis (TB) diagnosis often requires invasive procedures such as pleural biopsy. The aim of this study was to evaluate the role of real-time polymerase chain reaction (PCR) for the IS 6110 sequence of M. tuberculosis in pleural fluid specimens as a rapid and non-invasive test for pleural TB diagnosis. For this cross-sectional study, 150 consecutive patients with pleural effusion diagnosed by chest radiography, who were referred for diagnostic thoracocentesis and pleural biopsy and met eligibility criteria, had a pleural fluid specimen submitted for real-time PCR testing. Overall, 98 patients had pleural TB and 52 had pleural effusion secondary to other disease. TB diagnosis was obtained using acid-fast bacilli (AFB) smear or culture for mycobacteria and/or histopathologic examination in 94 cases and by clinical findings in 4 cases. Sensitivity, specificity, positive and negative predictive values of PCR testing for pleural TB diagnosis were 42.8% (95% CI 38.4 - 44.8), 94.2% (95% CI 85.8 - 98.0), 93.3% (95% CI 83.6 - 97.7), and 48.5% (95% CI 44.2 - 50.4), respectively. The real-time PCR test improved TB detection from 30.6% to 42.9% when compared to AFB smear and culture methods performed on pleural fluid specimens, although the best sensitivity was achieved by combining the results of culture and histopathology of pleural tissue specimens. The real-time PCR test of pleural fluid specimens is a useful and non-invasive additional assay for fast diagnosis of pleural TB.
Tài liệu tham khảo
Berger HW, Mejia E: Tuberculous pleurisy. Chest. 1973, 63: 88-92. 10.1378/chest.63.1.88.
Nance KV, Shermer RW, Askin FB: Diagnostic efficacy of pleural biopsy as compared with that of pleural fluid examination. Mod Pathol. 1991, 4: 320-324.
Valdes L, lvarez DA, San Jose E, Penela P, Valle JM, Garcıa-Pazos JM, Suarez J, Pose A: Tuberculous Pleurisy. A Study of 254 Patients. Arch Intern Med. 1998, 158: 2017-2021. 10.1001/archinte.158.18.2017.
Gopi A, Madhavan S, Sharma SK: Diagnosis and treatment of tuberculous pleural effusion in 2006. Chest. 2007, 131: 880-889. 10.1378/chest.06-2063.
Light RW: Update on tuberculous pleural effusion. Respirology. 2010, 15: 451-458. 10.1111/j.1440-1843.2010.01723.x.
Diacon AH, Van de Wal BW, Wyser C, Smedema JP, Bezuidenhout J, Bolliger CT, Walzl G: Diagnostic tools in tuberculous pleurisy: a direct comparative study. Eur Respir J. 2003, 22: 589-591. 10.1183/09031936.03.00017103a.
Centers for Disease Control and Prevention: Updated Guidelines for the Use of Nucleic Acid Amplification Tests in the Diagnosis of Tuberculosis. MMWR. 2009, 58: 7-10.
Pai M, Flores LL, Hubbard A, Riley LW, Colford JM: Nucleic acid amplification tets in the diagnosis of tuberculous pleuritis: a systematic review and meta-analysis. BMC Infect Dis. 2004, 4: 6-20. 10.1186/1471-2334-4-6.
Espy MJ, Uhl JR, Sloan LM, Buckwalter SP, Jones MF, Vetter EA, Yao JD, Wengenack NL, Rosenblatt JE, Cockerill FR, Smith TF: Real-time PCR in clinical microbiology: applications for routine laboratory testing. Clin Microbiol Rev. 2006, 19: 165-256. 10.1128/CMR.19.1.165-256.2006.
Morisson P, Neves DD: Evaluation of adenosine deaminase in the diagnosis of pleural tuberculosis: a Brazilian meta-analysis. J Bras Pneumol. 2008, 34: 217-224.
Seiscento M, Vargas FS, Rujula MJP, Bombarda S, Uip DE, Galesi VMN: Epidemiological aspects of pleural tuberculosis in the state of São Paulo, Brazil (1998-2005). J Bras Pneumol. 2009, 35: 548-554. 10.1590/S1806-37132009000600008.
Kent PT, Kubica GP: Public health mycobacteriology: a guide for level III laboratory. Atlanta: Centers of Disease Control Publication. 1985, N° PB 86-216-546
Boom R, Sol CJA, Salimans MMM, Jansen CL, Dillen PMEW, Noordaa JV: Rapid and simple method for purification of nucleic acids. J Clin Microb. 1990, 28: 495-503.
Hermans PW, van Sooligen D, Dale JW, Schuitema AR, McAdam RA, Catty D, van Embden JD: Insertion element IS986 from Mycobacterium tuberculosis: a useful tool for diagnosis and epidemiology of tuberculosis. J Clin Microb. 1990, 28: 2051-2058.
Verza M, Maschmann RA, Silva MS, Dalla Costa ER, Ribeiro MO, Rosso F, Suffys PN, Tortoli E, Marcelli F, Zaha A, Rossetti MRL: In house colorimetric reverse hybridisation assay for detection of the mutation most frequently associated with resistance to isoniazid in Mycobacterium tuberculosis. Mem Inst Oswaldo Cruz. 2009, 104: 710-714. 10.1590/S0074-02762009000500008.
Michelon CT, Rosso F, Schmid KB, Sperhacke RD, Oliveira MM, Kritski AL, Rezende L, Costa ERD, Ribeiro AW, Verza M, Cafrune PI, Nunes MS, Kuhleis D, Zaha A, Rossetti MLR: Colorimetric microwell plate reverse-hybridization assay for Mycobacterium tuberculosis detection. Memórias do Instituto Oswaldo Cruz. 2011, 106 (2): 194-199.
Cortez-Herrera E, Sperhacke RD, Becker D, Kritski A, Zaha A, Rossetti MLR: Internal control in PCR for Mycobacterium tuberculosis: usefulness and improvement of the diagnosis. Braz Arch Biol Technol. 2008, 51: 485-491. 10.1590/S1516-89132008000400006.
Nagesh BS, Sehgal S, Jindal SK, Arora SK: Evaluation of polymerase chain reaction for detection of Mycobacterium tuberculosis in pleural fluid. Chest. 2001, 119: 1737-1741. 10.1378/chest.119.6.1737.
Baba K, Pathak S, Sviland L, Langeland N, Hoosen AA, Asjo B, Dyrhol-Riise AM, Mustafa T: Real-time quantitative PCR in the diagnosis of tuberculosis in formalin-fixed paraffin-embedded pleural tissue in patients from a high HIV endemic area. Diagn Mol Pathol. 2008, 17: 112-117. 10.1097/PDM.0b013e31814ceac3.
Hillemann D, Galle J, Vollmer E, Richter E: Real-time PCR assay for improved detection of Mycobacterium tuberculosis complex in paraffin-embedded tissues. Int J Tuberc Lung Dis. 2006, 10: 340-342.
Moon JW, Chang Y S, Kim SK, Kim YS, Lee HM, Kim SK, Chang J: The clinical utility of polymerase chain reaction for the diagnosis of pleural tuberculosis. Clin Infect Dis. 2005, 41: 660-666. 10.1086/432474.
Gunisha P, Madhavan HN, Jayanthi U, Therese KL: Polymerase chain reaction using IS6110 primer to detect Mycobacterium tuberculosis in clinical samples. Indian J Pathol Microbiol. 2001, 44: 97-102.
Villena V, Rebollo MJ, Aguado JM, Galan A, Lopez Encuentra A, Palenque E: Polymerase chain reaction for the diagnosis of pleural tuberculosis in immunocompromised and immunocompetent patients. Clin Infect Dis. 1998, 26: 212-214. 10.1086/517025.
Lima D, Colares JK, Fonseca BAL: Combined use of the polymerase chain reaction and detection of adenosine deaminase activity on pleural fluid improves the rate of diagnosis of pleural tuberculosis. Chest. 2003, 124: 909-914. 10.1378/chest.124.3.909.
Chakravorty S, Sen MK, Tyagi JS: Diagnosis of extrapulmonary tuberculosis by smear, culture, and PCR using universal sample processing technology. J Clin Microbiol. 2005, 43: 4357-4362. 10.1128/JCM.43.9.4357-4362.2005.