Nested polymerase chain reaction for Mycobacterium tuberculosis IS6110 sequence on formalin-fixed paraffin-embedded tissues with granulomatous diseases for rapid diagnosis of tuberculosis

Luca Vago, Massimo Barberis, Andrea Gori, Paolo Scarpellini, Elena Sala, Manuela Nebuloni, Sara Bonetto, Maria Cannone, Giulia Marchetti, Fabio Franzetti, Giulio Costanzi

Research output: Contribution to journalArticlepeer-review

Abstract

We evaluated the sensitivity and specificity of a nested polymerase chain reaction (PCR) to the Mycobacterium tuberculosis IS6110 sequence on formalin-fixed paraffin-embedded tissue samples from patients with tubercular and other granulomatous lesions. Five groups of patients and samples were studied: (1) 28 samples from HIV-positive patients with tuberculosis, (2) 8 samples from HIV-negative patients with histologically suspected tuberculosis (confirmed by culture in 5 cases), (3) lymph nodes from 5 HIV-positive patients with Mycobacterium avium-intracellulare infection, (4) lymph nodes from 30 patients with sarcoidosis, and (5) specimens from 17 patients with other granulomatous diseases. The DNA was extracted from sections with a total thickness of 60 μm, and PCR amplified an internal fragment of 123 base pairs. All of the cases with M tuberculosis infection were PCR-positive, although this sensitivity was partially related to the initial concentration of the DNA used for amplification. Two of the group 4 samples also were repeatedly positive, thus reducing the specificity of the method. All of the cases with granulomatous diseases other than sarcoidosis were negative. We propose a simplified and highly sensitive nested PCR for the diagnosis of M tuberculosis infection on archived material in HIV-positive and HIV-negative patients.

Original languageEnglish
Pages (from-to)411-415
Number of pages5
JournalAmerican Journal of Clinical Pathology
Volume109
Issue number4
Publication statusPublished - 1998

Keywords

  • AIDS
  • HIV
  • Polymerase chain reaction [PCR]
  • Sarcoidosis
  • Tuberculosis

ASJC Scopus subject areas

  • Pathology and Forensic Medicine

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