title: Diagnostic value of a nested polymerase chain reaction assay on peripheral blood mononuclear cells from patients with pulmonary and extra-pulmonary tuberculosis.
abstract: METHODS
  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.
  
  
  METHODS
  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%).
  
  
  CONCLUSIONS
  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.
authors.affiliation: Service de Microbiologie, Hopital Saint Louis, Paris, France.
authors.forename: S
authors.initials: S
authors.lastname: Honore
authors.forename: J P
authors.initials: JP
authors.lastname: Vincensini
authors.forename: L
authors.initials: L
authors.lastname: Hocqueloux
authors.forename: M E
authors.initials: ME
authors.lastname: Noguera
authors.forename: D
authors.initials: D
authors.lastname: Farge
authors.forename: P
authors.initials: P
authors.lastname: Lagrange
authors.forename: J L
authors.initials: JL
authors.lastname: Herrmann
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D016022:Case-Control Studies; D003470:Culture Media; D004269:DNA, Bacterial / Q000235:genetics*; D005260:Female; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000382:microbiology*; D008297:Male; D008875:Middle Aged; D009169:Mycobacterium tuberculosis / Q000235:genetics* / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction; D011446:Prospective Studies; D012680:Sensitivity and Specificity; D013997:Time Factors; D014376:Tuberculosis / Q000097:blood* / Q000235:genetics*
chemical_list: D003470:Culture Media; D004269:DNA, Bacterial