title: [Study on rapid detecting of live Pseudomonas aeruginosa by EMA-PCR].
abstract: OBJECTIVE
  To establish a effective and rapid method by Ethidium Monoazide Bromide(EMA) in combination with PCR(EMA-PCR) for thedetection of live Pseudomrnonas aeruginosa.
  
  
  METHODS
  The oprI gene was used as the target gene for PCR detection of Pseudomonas aeruginosa , and PCR amplification was carried out by utilizing its pure isolates as the template. Sensitivity, EMA concentration and exposure time were optimized.
  
  
  RESULTS
  The sensitivity of PCR detection was 3 X 10(3) CFU/mIL, exposure time was 10 min. when the EMA concentration was not more than 5 μg/mLI, no obvious inhibition to the amplification of DNA derived from viable bacteria was observed. The PCR amplification of DNA derived from 3 X 10(6) CFU/mL dead cells could be inhibited effectively by EMA at the final concentration of 1 μg/mL. The results demonstrated the establised method could detect 1% live bacteria from a mixed bacterial population.
  
  
  CONCLUSIONS
  EMA-PCR can detect live bacteria of Pseudomonas aeruginosa effectively and avoid false positive result of the PCR detection.
authors.affiliation: Kunshan Center of Disease Control and Prevention of Jiangsu Province, Kunshan, China
authors.lastname: Fang Mei
authors.lastname: Lu Qiao-rong
authors.lastname: Wang Guo-qing
authors.lastname: Zhang Hong-bin
authors.lastname: Li Jian
mesh_terms: D001386:Azides; D015169:Colony Count, Microbial; D004269:DNA, Bacterial; D050296:Microbial Viability; D016133:Polymerase Chain Reaction / Q000379:methods*; D011550:Pseudomonas aeruginosa / Q000302:isolation & purification*
chemical_list: D001386:Azides; D004269:DNA, Bacterial; C014535:8-azidoethidium