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

Am J Clin Pathol. 1998 Apr;109(4):411-5. doi: 10.1093/ajcp/109.4.411.


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 microm, 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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • AIDS-Related Opportunistic Infections / diagnosis
  • Acquired Immunodeficiency Syndrome / complications
  • Biopsy
  • Evaluation Studies as Topic
  • Female
  • Formaldehyde
  • Granuloma / complications*
  • Humans
  • Mycobacterium avium-intracellulare Infection / complications
  • Mycobacterium avium-intracellulare Infection / diagnosis
  • Mycobacterium tuberculosis / isolation & purification*
  • Paraffin Embedding
  • Polymerase Chain Reaction
  • Sarcoidosis / complications
  • Sensitivity and Specificity
  • Tuberculosis / complications
  • Tuberculosis / diagnosis*


  • Formaldehyde