title: [EMA-pCR detection of enterohemorrhagic Eschrichia coli O157:H7].
abstract: OBJECTIVE
  A rapid and effective method with ethidium monoazide bromide (EMA) in combination with PCR (EMA-PCR) was established to detect live Enterohemorrhagic Eschrichia Coli O157:H7.
  
  
  METHODS
  The rfbE gene was used as the target gene for PCR detection of Eschrichia Coli O157:H7 by utilizing its pure isolates after the treatment of EMA as the template. The EMA concentration and reaction time was optimized.
  
  
  RESULTS
  The use of 10 microg/mL or less EMA did not inhibit the PCR amplification of DNA derived from viable bacteria. The PCR amplification of DNA derived from 2 x 10(7) CFU/mL dead cells can be inhibited by 0.5 microg/mL EMA. The sensitivity of the method was 2 x 10(4) CFU/mL. The results demonstrated that it could detect 1% live bacteria from a mixed bacterial population.
  
  
  CONCLUSIONS
  EMA-PCR can effectively detect live bacteria of O157:H7, it could be a potential rapid detection method applied in public health emergent events.
authors: West China School of Public Health, Sichuan University, Chengdu 610041, China.
authors: Ruo-Ying
authors: RY
authors: Huang
authors: Kunshan Center For Disease Control and Prevention, Kunshan 215300, China.
authors: Mei
authors: M
authors: Fang
authors: West China School of Public Health, Sichuan University, Chengdu 610041, China.
authors: Ying-Shu
authors: YS
authors: Luo
authors: Kunshan Center For Disease Control and Prevention, Kunshan 215300, China.
authors: Heng-Chuan
authors: HC
authors: Liu
authors: Kunshan Center For Disease Control and Prevention, Kunshan 215300, China.
authors: Qiao-Rong
authors: QR
authors: Lu
authors: West China School of Public Health, Sichuan University, Chengdu 610041, China.
authors: Guo-Qing
authors: GQ
authors: Wang
mesh_terms: D001386:Azides; D004269:DNA, Bacterial / Q000302:isolation & purification*; D019453:Escherichia coli O157 / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods*
chemical_list: D001386:Azides; D004269:DNA, Bacterial; C014535:8-azidoethidium