title: Restriction enzyme analysis of the hsp65 gene in clinical isolates from patients suspected of having pulmonary tuberculosis in Teresina, Brazil.
abstract: OBJECTIVE
  To identify mycobacterial species in the sputum of patients suspected of having pulmonary tuberculosis and to determine the impact that the acquisition of this knowledge has on the therapeutic approach.
  
  
  METHODS
  We evaluated 106 patients suspected of having pulmonary tuberculosis and referred to the pulmonology department of a public hospital in the city of Teresina, Brazil. Morning sputum specimens were evaluated for the presence of mycobacteria by sputum smear microscopy and culture. We used PCR and restriction enzyme analysis of the hsp65 gene (PRA-hsp65) to identify the strains of mycobacteria isolated in culture.
  
  
  RESULTS
  A total of 206 sputum samples were analyzed. Patient ages ranged from 15 to 87 years, and 67% were male. There was cough in 100% of the cases. The predominant radiographic pattern was moderate disease, observed in 70%. Smear positivity was 76%, and isolation in culture occurred in 91% of the cultures. Traditional tests identified nontuberculous mycobacteria (NTM) in 9% of the isolates. The PRA-hsp65 method confirmed these data, showing seven band patterns that were able to identify the isolated species of NTM: Mycobacterium kansasii; M. abscessus 1; M. abscessus 2; M. smegmatis; M. flavescens 1; M. gordonae 5; and M. gordonae 7. All of the patients with NTM were over 60 years of age, and bronchiectasis was seen in 88% of the X-rays. There were two cases of reinfection, initially attributed to M. abscessus and M. kansasii.
  
  
  CONCLUSIONS
  In immunocompetent patients, NTM can infect the lungs. It is important to identify the specific NTM in order to establish the correct diagnosis and choose the most appropriate therapeutic regimen. The PRA-hsp65 method is useful in identifying NTM species and can be implemented in molecular biology laboratories that do not specialize in the identification of mycobacteria.
authors: Universidade Federal do Piauí, Teresina, PI, Brasil.
authors: Maria das Graças Motta E
authors: Md
authors: Bona
authors: Maria José Soares
authors: MJ
authors: Leal
authors: Liline Maria Soares
authors: LM
authors: Martins
authors: Raimundo Nonato da
authors: RN
authors: Silva
authors: José Adail Fonseca de
authors: JA
authors: Castro
authors: Semiramis Jamil Hadad do
authors: SJ
authors: Monte
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001426:Bacterial Proteins / Q000235:genetics / Q000302:isolation & purification*; D001938:Brazil; D018834:Chaperonin 60 / Q000235:genetics / Q000302:isolation & purification*; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D009170:Nontuberculous Mycobacteria / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D015183:Restriction Mapping / Q000379:methods*; D013183:Sputum / Q000382:microbiology*; D014397:Tuberculosis, Pulmonary / Q000382:microbiology*; D055815:Young Adult
chemical_list: D001426:Bacterial Proteins; D018834:Chaperonin 60; C074119:heat-shock protein 65, Mycobacterium