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Detection of Mycobacterium tuberculosis in Paraffin-Embedded Pleural Biopsy Specimens by Commercial Ribosomal RNA and DNA Amplification Kits^*

^* From the Departments of Pneumology (Drs. Ruiz-Manzano, Monsó, and Martínez), Microbiology (Mr. Manterola, and Drs. Gamboa and Ausina), and Pathology (Dr. Calatrava), Hospital Universitari Germans Trias i Pujol, Badalona, Barcelona, Spain.

Correspondence to: Juan Ruiz-Manzano, MD, Pneumology Department, Hospital Universitari Germans Trias i Pujol, Carretera del Canyet s/n, 08916, Badalona, Barcelona, Spain; e-mail: jruiz{at}ns.hugtip.scs.es

Study objectives: To evaluate the utility of two gene amplification systems in historical paraffin-embedded pleural biopsy (PEB) tissues from patients with pleural tuberculosis, and to compare the results to those obtained with conventional histologic and microbiological methods.

Design: A retrospective study.

Patients and methods: Seventy-four formalin-fixed PEB tissues collected and stored over 12 years (1984 through 1995) were retrieved. Gene amplifications were performed in 57 tissues from patients with diagnoses of pleural tuberculosis and in 17 from patients with carcinoma as controls, using the first version of the Amplified Mycobacterium tuberculosis Direct Test (AMTDT; Gen-Probe; San Diego, CA) and the LCx Mycobacterium tuberculosis Assay (LCxMTB; Abbott Laboratories; Abbott Park, IL).

Results: The sensitivities of the AMTDT and LCxMTB were 52.6% and 63.2%, respectively (p = not statistically significant). The specificity of both tests was 100%. Twenty tissue samples (35.1%) were positive by both systems, and 10 tissues (17.5%) were positive only by the AMTDT, while 16 tissues (28.1%) were positive only by the LCxMTB. Both tests gave negative results for 11 specimens (19.3%). When both tests were used, a positive diagnosis was achieved in 80.7% of the samples. Diagnosis of 73.7% of patient conditions had previously been made by smear examination of pleural biopsy and sputum, pleural liquid, or biopsy culture. The overall diagnostic yield with both culture and amplification techniques was 96.5% (55 of 57 patients) for pleural tuberculosis, with amplification techniques adding 22.8% of the diagnoses.

Conclusions: Amplification techniques are useful in archival PEB tissues, providing additional diagnoses beyond culturing, although the sensitivity should be improved, possibly by standardizing protocols.

Key Words: gene amplification • paraffin-embedded pleural tissue • pleural tuberculosis

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