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Updated: Apr 18, 2026
Detection of Live Escherichia coli O157:H7 Cells by PMA-qPCR
Published on: February 1, 2014
[Study on rapid detecting of live Pseudomonas aeruginosa by EMA-PCR]
Fang Mei1, Lu Qiao-rong, Wang Guo-qing
1Kunshan Center of Disease Control and Prevention of Jiangsu Province, Kunshan, China
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.
Conclusion:
EMA-PCR can detect live bacteria of Pseudomonas aeruginosa effectively and avoid false positive result of the PCR detection.
