title: Rapid detection of Mycobacterium tuberculosis in respiratory specimens, blood and other non-respiratory specimens by amplification of rRNA.
abstract: METHODS
  Diagnostic methods employing gene technology based on amplification of DNA or RNA are expected to improve the speed, sensitivity, and specificity of Mycobacterium tuberculosis detection. The Amplified Mycobacterium Tuberculosis Direct Test (AMTDT) enables the amplification and detection of M. tuberculosis rRNA directly from respiratory specimens.
  
  
  OBJECTIVE
  To evaluate the performance of the AMTDT in direct detection of M. tuberculosis in respiratory specimens, blood and other clinical samples, and to compare this method with conventional culture and staining techniques.
  
  
  METHODS
  A total of 554 samples from 450 patients were examined in this study. All clinical specimens (with the exception of bone marrow aspirates and blood samples) were digested and decontaminated with sodium dodecyl (lauryl) sulfate (SDS)-NaOH. Bone marrow aspirates and blood samples were treated with 10% SDS. All processed samples were stained by auramine-rhodamine fluorochrome and inoculated onto Löwenstein-Jensen and Coletsos solid media, and into BACTEC-12B medium. In addition, the blood samples were inoculated into BACTEC 13A medium. The AMTDT was performed according to manufacturer's instructions. In those cases where discrepant results were obtained for AMTDT and cultures, patients' clinical data and other microbiological results were evaluated.
  
  
  RESULTS
  The sensitivity, specificity, and positive and negative predictive values for AMTDT were 87.5, 100, 100, and 96.7%, respectively, in respiratory specimens and 86.8, 100, 100, and 92.8%, respectively, in non-respiratory specimens. The differences in sensitivity of these two groups of specimens were not highly statistically significant (P > 0.005).
  
  
  CONCLUSIONS
  The sensitivity and specificity of the AMTDT were satisfactory for detection of M. tuberculosis in all types of clinical samples. Some minor changes in assay format and laboratory protocols may increase the sensitivity of the AMTDT without adversely affecting its specificity.
authors.affiliation: Servicio de Microbiología, Hospital Universitario Germans Trias i Pujol, Barcelona, Spain.
authors.forename: F
authors.initials: F
authors.lastname: Gamboa
authors.forename: J M
authors.initials: JM
authors.lastname: Manterola
authors.forename: J
authors.initials: J
authors.lastname: Lonca
authors.forename: B
authors.initials: B
authors.lastname: Viñado
authors.forename: L
authors.initials: L
authors.lastname: Matas
authors.forename: M
authors.initials: M
authors.lastname: Giménez
authors.forename: J R
authors.initials: JR
authors.lastname: Manzano
authors.forename: C
authors.initials: C
authors.lastname: Rodrigo
authors.forename: P J
authors.initials: PJ
authors.lastname: Cardona
authors.forename: E
authors.initials: E
authors.lastname: Padilla
authors.forename: J
authors.initials: J
authors.lastname: Dominguez
authors.forename: V
authors.initials: V
authors.lastname: Ausina
mesh_terms: D001431:Bacteriological Techniques; D001769:Blood / Q000382:microbiology; D001992:Bronchoalveolar Lavage Fluid / Q000382:microbiology; D005243:Feces / Q000382:microbiology; D005784:Gene Amplification; D006801:Humans; D008198:Lymph Nodes / Q000382:microbiology; D015336:Molecular Probe Techniques; D009169:Mycobacterium tuberculosis / Q000235:genetics / Q000302:isolation & purification*; D011237:Predictive Value of Tests; D012329:RNA, Bacterial / Q000235:genetics*; D012680:Sensitivity and Specificity; D013183:Sputum / Q000382:microbiology; D014556:Urine / Q000382:microbiology; D012407:rRNA Operon / Q000235:genetics
chemical_list: D012329:RNA, Bacterial