Diagnostic value of a nested polymerase chain reaction assay on peripheral blood mononuclear cells from patients with pulmonary and extra-pulmonary tuberculosis

• Published in: The international journal of tuberculosis and lung disease Vol. 5; no. 8; pp. 754 - 762
• Main Authors: HONORE, S, VINCENSINI, J. P, HOCQUELOUX, L, NOGUERA, M. E, FARGE, D, LAGRANGE, P, HERRMANN, J. L
• Format: Journal Article
• Language: English
• Published: Paris The International Union Against Tuberculosis & Lung Disease 01.08.2001; Union internationale contre la tuberculose et les maladies respiratoires
• Subjects: Adult; Aged; Aged, 80 and over; Bacterial diseases; Biological and medical sciences; Case-Control Studies; Culture Media; Diagnostic; DNA, Bacterial - genetics; Female; Human bacterial diseases; Humans; Infectious diseases; Leukocytes, Mononuclear - microbiology; Male; Medical sciences; Middle Aged; Mycobacterium tuberculosis - genetics; Mycobacterium tuberculosis - isolation & purification; nested PCR; Polymerase Chain Reaction; Prospective Studies; Sensitivity and Specificity; Time Factors; Tuberculosis; Tuberculosis - blood; Tuberculosis - genetics; Tuberculosis and atypical mycobacterial infections; Human; Lung disease; Nested polymerase chain reaction; Respiratory disease; Lung; Mycobacterial infection; Blood; Infection; Mononuclear cell; Tuberculosis; Bacteriosis; Diagnosis; Molecular biology
• ISSN: 1027-3719

SETTING: Saint Louis Hospital, Paris, France.OBJECTIVE: To determine the clinical relevance of detection of Mycobacterium tuberculosis DNA by nested polymerase chain reaction (PCR) in peripheral blood mononuclear cells (PBMCs) in the rapid diagnosis of tuberculosis.DESIGN: Single-centre prospective case study of 90 hospitalised patients and 50 healthy subjects or blood donors from 1 January to 30 June 1998.RESULTS: Twenty-three patients were diagnosed with tuberculosis (26.7%); 20 tuberculosis patients were culture-positive, with seven smear-positive for acid-fast bacilli. Sensitivity of smear, culture and nested PCR was 30.4 (7/23), 87 (20/23) and 30.4% (7/23), respectively. The specificity of smear and culture was 100%, and the specificity of the nested PCR was 96% in the healthy subjects. However, the specificity decreased to 83.6% in the hospitalised patients, with 11 nested PCR-positive patients without a diagnosis of tuberculosis. The sensitivity of the nested PCR was low in pulmonary tuberculosis (22.2%), but increased in pulmonary/extra-pulmonary tuberculosis (50%), extra-pulmonary tuberculosis (33%), and disseminated tuberculosis (33%).CONCLUSION: The use of a nested PCR assay on PBMC may pose problems for the rapid diagnosis of tuberculosis with regard to low sensitivity and specificity. However, further studies are needed to confirm this technique as an alternative test for the diagnosis of paucibacillary forms of tuberculosis.