Amir Hossein Jafarian - Department of Pathology, Ghaem Hospital, Ahmadabad Blvd., Mashhad University of Medical Sciences (MUMS) Mashhad, Iran
Abbasali Omidi - Department of Pathology, Ghaem Hospital, Ahmadabad Blvd., Mashhad University of Medical Sciences (MUMS) Mashhad, Iran
Javad Ghenaat - Department of Microbiology, Ghaem Hospital, Ahmadabad Blvd., MUMS, Mashhad, Iran
Kiarash Ghazvini - Department of Microbiology, Ghaem Hospital, Ahmadabad Blvd., MUMS, Mashhad, Iran
Hossein Ayatollahi - Department of Biochemistry, Ghaem Hospital, Ahmadabad Blvd., MUMS, Mashhad, Iran
Minoo Erfanian - Department of Pathology, Ghaem Hospital, Ahmadabad Blvd., Mashhad University of Medical Sciences (MUMS) Mashhad, Iran
Mohammadd Taghi Shakeri - Department of Microbiology, Ghaem Hospital, Ahmadabad Blvd., MUMS, Mashhad, Iran
Mahmoud Bagheri - Department of Microbiology, Ghaem Hospital, Ahmadabad Blvd., MUMS, Mashhad, Iran
Hooman Tavassolian - Department of Pathology, Ghaem Hospital, Ahmadabad Blvd., Mashhad University of Medical Sciences (MUMS) Mashhad, Iran

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 ۱۰ cases with bronchogenic carcinoma as controls were investigated. We designed a Multiplex PCR DNA amplification method with two targets: ۱۲۳bp DNA fragment from IS۶۱۱۰, which is present only in mycobacterium tuberculosis complex and ۱۶۲bp DNA encoding Ag ۸۵complex which is present in all of mycobacteria. The FFPE also stained by Acid fast and Rhodamine-Auramine   staining method. Results In ۲۶ samples (۶۵%) ۱۲۳ bp and ۱۶۲ bp DNA fragments were detected together (۱۲ in bronchial samples and ۱۴ in pleural samples).The ۱۶۲ bp fragment wasn't detected alone. The sensitivity of PCR was ۶۵% and the specificity was ۱۰۰%. Eleven cases were positive for Acid fast staining. There was ۲۷.۵% sensitivity and ۱۰۰% specificity. Thirteen cases were positive for Auramine-Rhodamine staining (A-R-S); there was ۳۲.۵% sensitivity and ۱۰۰% specificity. All of the ۱۰ controls were negative for ۱۲۳ bp, ۱۶۲ 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.

Acid fast stain, Auramine-Rhodamin stain, Bronchial tissue, M. tuberculosis, Multiplex PCR, Pleural tissue