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Propidium monoazide–quantitative polymerase chain reaction (PMA-qPCR) assay for rapid detection of viable and viable but non-culturable (VBNC) Pseudomonas aeruginosa in swimming pools

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Abstract

Lack of culturability in the viable but non-culturable (VBNC) bacteria and the ability to regain infectivity in favourable conditions is one of the new challenges of public health providers for Pseudomonas aeruginosa monitoring in environmental samples. Propidium monoazide quantitative polymerase chain reaction (PMA-qPCR) is one of the promising methods for timely detection of VBNC pathogens in environmental samples. We developed and used a method for the first time to detection of VBNC P. aeruginosa in swimming pool water samples using a membrane filter (MF). Moreover, the dominant model of the distribution of colonies on the MF and the effect of the culture medium and MF type on colony recovery by MF were evaluated. Swimming pool samples were subjected to conventional culture-based, qPCR and PMA-qPCR methods and the results were compared for the presence of VBNC P. aeruginosa in the samples. The positivity rate was 21% and 75% for P. aeruginosa in water samples as confirmed by standard culture-based and qPCR methods, respectively. Furthermore, of 24 samples, 9 (37.5%) were positive for VBNC P. aeruginosa. The developed qPCR/PMA-qPCR assay can detect the VBNC bacteria directly from aquatic samples and may result in better monitoring of recreational waters.

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Acknowledgements

This study is a part of PhD thesis conducted by the first author submitted to Tehran University of Medical Sciences. The authors acknowledge the Center for Water Quality Research (CWQR), Institute for Environmental Research (IER) and Tehran University of Medical Sciences (TUMS), Tehran, Iran, for financially supporting this project (Project No: 96-01-46-34273).

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Authors and Affiliations

  1. Department of Environmental Health Engineering, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran

    Abdolali Golpayegani, Mahmood Alimohammadi & Ramin Nabizadeh Nodehi

  2. Vice-Chancellor for Health, Bam University of Medical Sciences, Bam, Iran

    Abdolali Golpayegani

  3. Environmental Health Engineering Research Center, Kerman University of Medical Sciences, Kerman, Iran

    Abdolali Golpayegani

  4. Division of Microbiology, Department of Pathobiology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran

    Masoumeh Douraghi

  5. Department of Virology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran

    Farhad Rezaei

  6. Center for Water Quality Research (CWQR), Institute for Environmental Research (IER), Tehran University of Medical Sciences, PO Box 14155-6446, Tehran, Iran

    Mahmood Alimohammadi & Ramin Nabizadeh Nodehi

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  1. Abdolali Golpayegani
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  2. Masoumeh Douraghi
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  4. Mahmood Alimohammadi
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Correspondence to Ramin Nabizadeh Nodehi.

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Golpayegani, A., Douraghi, M., Rezaei, F. et al. Propidium monoazide–quantitative polymerase chain reaction (PMA-qPCR) assay for rapid detection of viable and viable but non-culturable (VBNC) Pseudomonas aeruginosa in swimming pools. J Environ Health Sci Engineer 17, 407–416 (2019). https://doi.org/10.1007/s40201-019-00359-w

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  • DOI: https://doi.org/10.1007/s40201-019-00359-w

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