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Updated: May 3, 2026
Detection of Live Escherichia coli O157:H7 Cells by PMA-qPCR
Published on: February 1, 2014
[EMA-pCR detection of enterohemorrhagic Eschrichia coli O157:H7]
Ruo-Ying Huang1, Mei Fang2, Ying-Shu Luo1
1West China School of Public Health, Sichuan University, Chengdu 610041, China.
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.
Conclusion:
EMA-PCR can effectively detect live bacteria of O157:H7, it could be a potential rapid detection method applied in public health emergent events.
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