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

Publish Year: 1387
نوع سند: مقاله ژورنالی
زبان: English
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شناسه ملی سند علمی:

JR_IJBMS-10-4_002

تاریخ نمایه سازی: 27 مهر 1400

Abstract:

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.

Authors

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

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