title: Incidence of tuberculous and non-tuberculous mycobacteria, differentiated by multiplex PCR, in clinical specimens of a large general hospital.
abstract: OBJECTIVE
  To determine the incidence of Mycobacterium tuberculosis complex and non-tuberculous mycobacterial isolates in the routine setting of a large general hospital using an "in-house" multiplex polymerase chain reaction method and to establish a paradigm for the definitive identification of mycobacteria isolated using semi-automated equipment.
  
  
  METHODS
  Established tests, including polymerase chain reaction restriction enzyme analysis, PNB, and NAP inhibition tests as the gold standard, showed 100% agreement with an IS6110/hsp65 multiplex polymerase chain reaction when used to identify stock strains (n = 117).
  
  
  RESULTS
  In a subsequent study, 8,790 clinical specimens producing 476 isolates were evaluated with multiplex PCR and also showed 100% agreement in identification using PRA-polymerase chain reaction as the gold standard. The application of this technique to routine analysis was demonstrated in this study. A method was established with the initial application of multiplex PCR for all positive liquid cultures and the subsequent identification of non-tuberculous mycobacteria by polymerase chain reaction restriction enzyme analysis. In total, 77% of isolates belonged to the Mycobacterium tuberculosis complex, and 23% were non-tuberculous mycobacteria.
  
  
  CONCLUSIONS
  Several non-tuberculous mycobacterial species were identified, primarily M. avium, but other potentially pathogenic species were also frequently observed, including M. fortuitum, M. abscessus, and M. kansasii. The expeditious communication of these data to the clinical staff was fundamental for the diagnosis of clinical cases. Even in settings where tuberculosis is of major importance, the incidence of non-tuberculous mycobacteria infection is substantial.
authors.affiliation: Faculty of Medical Sciences, University of Campinas, Campinas, SP, Brazil.
authors.forename: Eliane Picoli Alves
authors.initials: EP
authors.lastname: Bensi
authors.forename: Patricia Costa
authors.initials: PC
authors.lastname: Panunto
authors.forename: Marcelo de Carvalho
authors.initials: Mde C
authors.lastname: Ramos
mesh_terms: D001938:Brazil; D006769:Hospitals, General; D006801:Humans; D060885:Multiplex Polymerase Chain Reaction; D009169:Mycobacterium tuberculosis / Q000145:classification / Q000302:isolation & purification*; D009170:Nontuberculous Mycobacteria / Q000145:classification / Q000302:isolation & purification*; D015183:Restriction Mapping