title: Nested polymerase chain reaction for Mycobacterium tuberculosis IS6110 sequence on formalin-fixed paraffin-embedded tissues with granulomatous diseases for rapid diagnosis of tuberculosis.
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 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.
authors.affiliation: Pathology Unit, L. Sacco Institute of Biomedical Sciences, L. Sacco Hospital, University of Milan, Italy.
authors.forename: L
authors.initials: L
authors.lastname: Vago
authors.forename: M
authors.initials: M
authors.lastname: Barberis
authors.forename: A
authors.initials: A
authors.lastname: Gori
authors.forename: P
authors.initials: P
authors.lastname: Scarpellini
authors.forename: E
authors.initials: E
authors.lastname: Sala
authors.forename: M
authors.initials: M
authors.lastname: Nebuloni
authors.forename: S
authors.initials: S
authors.lastname: Bonetto
authors.forename: M
authors.initials: M
authors.lastname: Cannone
authors.forename: G
authors.initials: G
authors.lastname: Marchetti
authors.forename: F
authors.initials: F
authors.lastname: Franzetti
authors.forename: G
authors.initials: G
authors.lastname: Costanzi
mesh_terms: D017088:AIDS-Related Opportunistic Infections / Q000175:diagnosis; D000163:Acquired Immunodeficiency Syndrome / Q000150:complications; D001706:Biopsy; D005069:Evaluation Studies as Topic; D005260:Female; D005557:Formaldehyde; D006099:Granuloma / Q000150:complications*; D006801:Humans; D015270:Mycobacterium avium-intracellulare Infection / Q000150:complications / Q000175:diagnosis; D009169:Mycobacterium tuberculosis / Q000302:isolation & purification*; D016612:Paraffin Embedding; D016133:Polymerase Chain Reaction; D012507:Sarcoidosis / Q000150:complications; D012680:Sensitivity and Specificity; D014376:Tuberculosis / Q000150:complications / Q000175:diagnosis*
chemical_list: D005557:Formaldehyde