Comparison of Multiplex PCR and Acid Fast and Auramine -Rhodamine Staining for Detection of Mycobacterium tuberculosis and Non tuberculosis Mycobacteria in Paraffin- Embedded Pleural and Bronchial Tissues with Granulomatous Inflammation and Caseous Necrosis | ||
| Iranian Journal of Basic Medical Sciences | ||
| مقاله 2، دوره 10، شماره 4، دی 2008، صفحه 216-221 اصل مقاله (198.96 K) | ||
| نوع مقاله: Original Article | ||
| شناسه دیجیتال (DOI): 10.22038/ijbms.2008.5233 | ||
| نویسندگان | ||
| Amir Hossein Jafarian1؛ Abbasali Omidi* 1؛ Javad Ghenaat2؛ Kiarash Ghazvini2؛ Hossein Ayatollahi3؛ Minoo Erfanian1؛ Mohammadd Taghi Shakeri2؛ Mahmoud Bagheri2؛ Hooman Tavassolian1 | ||
| 1Department of Pathology, Ghaem Hospital, Ahmadabad Blvd., Mashhad University of Medical Sciences (MUMS) Mashhad, Iran | ||
| 2Department of Microbiology, Ghaem Hospital, Ahmadabad Blvd., MUMS, Mashhad, Iran | ||
| 3Department of Biochemistry, Ghaem Hospital, Ahmadabad Blvd., MUMS, Mashhad, Iran | ||
| چکیده | ||
| Objective The aim of this study was to compare the sensitivity and specificity of Acid fast and Auramine-Rhodamine staining and Multiplex PCR for the detection of Mycobacterium tuberculosiscomplex and non tuberculosis Mycobacteria on formalin fixed paraffin embedded tissues (FFPE) Materials and Methods Forty cases of FFPE pleural and bronchial tissue with chronic granulomatous inflammation and caseous necrosis and 10 cases with bronchogenic carcinoma as controls were investigated. We designed a Multiplex PCR DNA amplification method with two targets: 123bp DNA fragment from IS6110, which is present only in mycobacterium tuberculosis complex and 162bp DNA encoding Ag 85complex which is present in all of mycobacteria. The FFPE also stained by Acid fast and Rhodamine-Auramine staining method. Results In 26 samples (65%) 123 bp and 162 bp DNA fragments were detected together (12 in bronchial samples and 14 in pleural samples).The 162 bp fragment wasn't detected alone. The sensitivity of PCR was 65% and the specificity was 100%. Eleven cases were positive for Acid fast staining. There was 27.5% sensitivity and 100% specificity. Thirteen cases were positive for Auramine-Rhodamine staining (A-R-S); there was 32.5% sensitivity and 100% specificity. All of the 10 controls were negative for 123 bp, 162 bp DNA fragments, for Acid fast and Auramine-Rhodamine staining. Conclusion Multiplex PCR is a sensitive, specific and rapid method for detection of M. tuberculosis in FFPE tissues. | ||
| کلیدواژهها | ||
| Acid fast stain؛ Auramine-Rhodamin stain؛ Bronchial tissue؛ M. tuberculosis؛ Multiplex PCR؛ Pleural tissue | ||
| مراجع | ||
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1.Chakravorty S, Sen MK, Tyagi JS. Diagnosis of Extrapulmonary tuberculosis by smear, culture, and PCR using universal sample processing technology. J Clin Microbiol 2005 Sep; 43 (9): 4357- 4362.
2.Li Z, W Qin W, Yue, Meng Q, Lin Q. Study on detection of the mycobacteria DNA in formalin-fixed, paraffin-embedded tissue samples by triplex polymerase chain reaction. Wei Sheng Wu Xue Bao 2002 Feb; 42(1): 69-75.
3.Fukunaga H, Murakami T,Gondo T,sugi K, Ishihara T. Sensitivity of Acid- fast staining for mycobacterium. Tuberculosis in formalin- fixed tissues. Am J Respir crit care med 2002; 166. 994-997.
4.Li JY, lo ST, Ng CS. Molecular detection of mycobacterium tuberculosis in tissues showing granulomatous inflamation without demonstrable acid fast bacilli. Diagn Mol pathol 2000; 9: 67-74.
5. 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.
6. Selva E, Hofman V, Berto F, Musso S, Castillo L. The value of polymerase chain reaction detection of Mycobacterium tuberculosis in granulomas isolated by laser capture microdissection. Pathology 2004 Feb; 36 (1): 77-81.
7. Mustafa AS, Abal AT, chugh TD. Detection of Mycobacterium tuberculosis complex and non tuberculous Mycobacteria by Multiplex polymerase chain reaction. East Medit Health J 1999; 5 (1): 61-70.
8. Wiker HG, Sletten K,Nagai S, Harboe M. Evidence for three separate genes encoding the proteins of the My cobacterial antigen 85 complex. Infect Immun 1990 Jan; 58(1): 272-4.
9.Sharma S, Mallick GP, Verma R, Ray SK. Polymerase chain reaction Amplification of IS6110 Sequence to detect Mycobacterium tuberculosis complex from formalin – fixed paraffin- embedded Tissues of Deer. Vet Res Commun 2007 Jan; 31 (1): 17-21.
10.Bass Jr JB,Laurece S,Philip C,Richard F,Dixie E.. Diagnostic standard and classification of tuberculosis and other Mycobacterial disease. Am review of respiratory disease 1999; 123: 343-58.
11.Van soolingen D,Hermans PW,Dehass PE,Vanembden JD. Insertion element IS1081-associated restriction fragment length polymorphism in Mycobacterium tuberculosis complex species: a reliable tool for recognizing Mycobacterium bovis BCG. J Clinic Microbiol 1992 Jul; 30(7): 1772-7.
12.Jambhekar NA, Kulkarni SP, Madur BP. Agarwal S, Rajan MG. Application of the polymerase chain reaction on formalin-fixed, paraffin embedded tissue in the recognition of tuberculous ostemyelitis. J Bone Joint surg Br 2006 Aug; 88 (8): 1097-101.
13.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 Mar; 10(3): 340-2.
14.Fite E, Fenandez Figueras MT, Prats R, Vaquero M, Morera J. High prevalence of Mycobacterium tuberculosis DNA in biopsies from sarcoidosis patients from Catalonia, spain. Respiration 2006; 73 (1): 14-5.
15.Ardeleanue C, Andrei F, ceausa M, Ene D, Mihai M, Butur G. et al. Cellurar immune response in atypical tuberculosis diagnosed by PCR in paraffin embedded material. Rom J Morphol EmbryoL 1999-2004; 45: 63-72.
16.Schultz S, Carbras AD, Kremer M, Weirich G, M Micthke T, et al. Species identification of Mycobacteria in paraffin embedded tissues: frequent detection of nontuberculous Mycobacteria. Mod Pathol 2005 Feb; 18(2): 274-82.
17.Zink AR, erlich AG. Molecular strain identification of the Mycobacterium tuberculosis complex in archival tissue samples. J Clin pathol. 2004 Nov; 57 (11): 1185-92.
18.Cataloluk O, Cakmak EA, Buyukberber N, Barlas O. Formalin Fixing and paraffin embedding may lead to extra band development in PCR. New Microbiol 2003 Apr; 26(2): 193-8.
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