Nested polymerase chain reaction for Mycobacterium tuberculosis IS6110 sequence on formalin-fixed paraffin-embedded tissues with granulomatous diseases for rapid diagnosis of tuberculosis.
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.
Vago
L
L
Pathology Unit, L. Sacco Institute of Biomedical Sciences, L. Sacco Hospital, University of Milan, Italy.
Barberis
M
M
Gori
A
A
Scarpellini
P
P
Sala
E
E
Nebuloni
M
M
Bonetto
S
S
Cannone
M
M
Marchetti
G
G
Franzetti
F
F
Costanzi
G
G
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*
D005557:Formaldehyde