title: A multiplex PCR method for detection of Aspergillus spp. and Mycobacterium tuberculosis in BAL specimens.
abstract: OBJECTIVE
  The aim of this study was the detection of Aspergillus species and Mycobacterium tuberculosis together in bronchoalveolar lavage (BAL) using of multiplex PCR.
  
  
  METHODS
  In this study, from September 2012 until June 2013, 100 bronchoalveolar lavage (BAL) specimens were collected from patients suspected of tuberculosis (TB). After the direct and culture test, multiplex PCR were utilized in order to diagnose Aspergillus species and M. tuberculosis. Phenol-chloroform manual method was used in order to extract DNA from these microorganisms. Aspergillus specific primers, M. tuberculosis designed primers and beta actin primers were used for multiplex PCR.
  
  
  RESULTS
  In this study, by multiplex PCR method, Aspergillus species were identified in 12 samples (12%), positive samples in direct and culture test were respectively 11% and 10%. Sensitivity and specificity of this method in comparison to direct test were respectively 100% and 98.8%, also sensitivity and specificity of this method in comparison to culture test were respectively 100% and 97.7%. In this assay, M. tuberculosis was identified in 8 samples (8%). Mycobacterium-positive samples in molecular method, direct and culture test were respectively 6%, 5% and 7%. Sensitivity and specificity of PCR method in comparison to direct test were 80% and 97.8% also sensitivity and specificity of this method in comparison to culture test was 71.4% and 98.9%.
  
  
  CONCLUSIONS
  In the present study, multiplex PCR method had higher sensitivity than direct and culture test in order to identify and detect Aspergillus, also this method had lower sensitivity for identification of M. tuberculosis, suggesting that the method of DNA extraction was not suitable.
authors.affiliation: Department of biology, Islamic Azad university, Varamin-Pishva branch, Tehran, Iran. Electronic address: farzaneh.amini@rocketmail.com.
authors.forename: F
authors.initials: F
authors.lastname: Amini
authors.affiliation: Molecular biology research center, Baqiyatallah university of medical sciences, Tehran, Iran. Electronic address: kachueiz@yahoo.com.
authors.forename: R
authors.initials: R
authors.lastname: Kachuei
authors.affiliation: Department of biology, Islamic Azad university, Varamin-Pishva branch, Tehran, Iran. Electronic address: niloofar_noorbakhsh@yahoo.com.
authors.forename: F
authors.initials: F
authors.lastname: Noorbakhsh
authors.affiliation: Applied microbiology research center, Baqiyatallah university of medical sciences, Tehran, Iran. Electronic address: imanifouladi.a@gmail.com.
authors.forename: A A
authors.initials: AA
authors.lastname: Imani Fooladi
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001230:Aspergillus / Q000235:genetics / Q000302:isolation & purification*; D001992:Bronchoalveolar Lavage Fluid / Q000382:microbiology*; D004271:DNA, Fungal / Q000032:analysis / Q000302:isolation & purification; D005260:Female; D006801:Humans; D008297:Male; D008828:Microbiological Techniques; D008875:Middle Aged; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D009169:Mycobacterium tuberculosis / Q000235:genetics / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D014376:Tuberculosis / Q000175:diagnosis / Q000382:microbiology
chemical_list: D004271:DNA, Fungal
keywords: Aspergillus; BAL; Mycobacterium tuberculosis; PCR