Document Type : Original Research
Authors
Molecular Pathology Research Center, Kermanshah University of Medical Sciences, Kermanshah, Iran
Abstract
Background and Objective: Tuberculosis is still a major health problem, involving about 1/3 of the world´s population. Diagnosis is difficult when we only use Ziehl-Neelson staining. Many cases may be missed. A more rapid and sensitive diagnostic method is necessary. PCR may be helpful. The aim of this study was to compare PCR, Zieh-Neelsen staining and histopathologic findings in diagnosis of tuberculosis on formalin-fixed paraffin-embedded tissues. Methods: Paraffin blocks of the submitted specimens of the patients clinically suspicious for tuberculosis or containing granuloma were selected. Ziehl-Neelsen Staining & TB-PCR (IS6110 element) was carried out. The results of the tests were compared by using the McNemar test. Statistical significance was accepted when the P value was less than 0.05. Results: Forty five specimens were included in the study, 35 had granulomas (19 with caseous necrosis). Acid-fast bacilli were identified in 17 specimens (37.8%). TB-PCR was positive in 16specimens (84%) with caseating granulomatous, 11 specimens (68.8%) with non-caseating granulomas & 6 specimens (60%) without granulomas. (P value = 0.59). Conclusions: TB-PCR on paraffin–embedded tissue is a potentially useful approach for early, rapid and sensitive diagnosis of tuberculosis. It is especially useful when granuloma is seen in tissue section, while acid-fast stain is negative. If there was no facilities for PCR, histopathological diagnosis with clinical correlation are more reliable in comparison to AFB results.
Keywords
- Bahadori M, Azizi MH. Common challenges in laboratory diagnosis and management of tuberculosis. Red Crescent Med J 2012;14(1):3-9.
- WHO.WHO report Global tuberculosis control- Surveillance, Planning and Financing. WHO Press, 2008: 22-3.
- WHO.WHO report Global tuberculosis control- Epidemiology, Strategy and Financing. WHO Press, 2009:10-6.
- Hemmati M, Seghatoleslam A, Rasti M, Ebadat S, Mosavari N, Habibagahi M, et al.Expression and purification of recombinant mycobacterium tuberculosis (TB) antigens, ESAT-6, CFP-10 and ESAT-6/CFP-10 and their diagnosis potential for detection of TB patients. Iran Red Crescent Med J 2011;13(8):556-63.
- Arora SK, Gupta V, Gupta A, Bambery P, Kapoor GS, Sehgal S. Diagnostic efficacy of polymerase chain reaction in granulomatous uveitis. Tuber Lung Dis 1999;79 (4):229-33.
- Jahansen IS, Thomsen VØ, Forsgren A, Hansen BF, and Lundgren B. Detection of Mycobacterium tuberculosis complex in formalin-embedded tissue specimens with necrotizing granulomatous inflammation by strand displacement amplification. J Mol Diagn 2004;6(3):231-6.
- Chakravorty S, Sen MK, Tyagi JS. Diagnosis of Extra pulmonary tuberculosis by smear, culture, and PCR using universal sample processing technology. J Clin Microbiol 2005;43(9):4357-62.
- Park DY, Kim JY, Choi KU, Lee JS, Lee CH, Sol MY, et al. Comparison of polymerase chain reaction with histopathologic eatures for diagnosis of tuberculosis in formalin-fixed, paraffin-embedded histologic specimens. Arch Pathol Lab Med 2003;127(3):326-30.
- Osaki M, Adachi H, Gomyo Y, Yoshida H, Ito H. Detection of mycobacterial DNA in formalin-fixed, paraffin-embedded tissue specimens by duplex polymerase chain reaction: application to histopathologic diagnosis. Mod Pathol 1997;10(1):78-83.
- Kivihya-Ndugga L, Van Cleeff M, Juma E, Kimwomi J, Githui W, Oskam L, et al. Comparison of PCR with the routine procedure for diagnosis of tuberculosis in a population with high prevalence of tuberculosis and human immunodeficiency virus. J Clin Microbiol 2004;42(3):1012-5.
- Tarng DC, Su WJ, Huang TP. PCR diagnosis on formalin-fixed, paraffin-embedded tissues with acid-fast stain and culture negative in chronic in chronic dialysis patients of cervico-mediastinal tubercles lymphadenitis. Nephrol Dial Transplant 1998;13(6):1543-6.
- Negi SS, Khan SF, Gupta S, Pasha ST, Khare S, Lal S. Comparison of the conventional diagnostic modalities, bactec culture and polymerase chain reaction test for diagnosis of tuberclosis. Indian J Med Microbiol 2005;23(1):29-33.
- Omidi AA,Ghenaat J,Ghazvini K,Ayatollahi H,tavassolian h,Jafarian AH, et al. Incidence of Mycobacterium tuberculosis detection in formalin fixed-paraffin embedded granulomatous dermatoses with multiplex PCR comparing fluorescent microscopy and acid fast staining in eastern Iran. Internet J Microb 2007;4(1):216-22.
- Bancroft J D, Stevens A, Turner D R. Theory and Practice of Histological Techniques. Churchill Livingston Inc, New York. Third edition 1990;112:294-296.
- Kumar R,Abbas A,Delancey A, Malone E. Pathologic Basis of Disease. 8th ed. Philadelphia: Saunders Elsevier;2010.
- Varello K, Pezzolato M, Mascarino D, Ingravalle F, Caramelli M, Bozzetta E. Comparison of histologic techniques for the diagnosis of bovine tuberculosis in the framework of eradication programs. J Vet Diagn Invest 2008;20(2):164-9.
- Salian NV, Rish JA, Eisenach KD, Cave MD, Bates JH. Polymerase chain reaction to detect Mycobacterium tuberculosis in histologic specimens. Am J Respir Crit Care Med 1998;158(4):1150-5.
- Nopvichai C, Sanpavat A, Sawatdee R, Assanasen T, Wacharapluesadee S, Thorne P S, et al.PCR detection of Mycobacterium tuberculosis in necrotizing non-granulomatous lymphadenitis using formalin-fixed paraffin-embedded tissue: a study in Thai patients. J Clin Pathol 2009;62(9):812-5.