Nested Polymerase Chain Reaction forMycobacterium tuberculosisIS6110 Sequence on Formalin-Fixed Paraffin-Embedded Tissues With Granulomatous Diseases for Rapid Diagnosis of Tuberculosis

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Abstract
We evaluated the sensitivity and specificity of a nested polymerase chain reaction (PCR) to the Mycobacterium tuberculosis IS6110 sequence on formalin-fixed paraffinembedded 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 HIVpositive patients with Mycobacterium avium-intraceliulare 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 urn, 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.

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