Rapid and Accurate Detection of Bacteriophage Activity against Escherichia coli O157:H7 by Propidium Monoazide Real-Time PCR
Joint Authors
Liu, Hui
Niu, Yan D.
Li, Jinquan
Stanford, Kim
McAllister, Tim A.
Source
Issue
Vol. 2014, Issue 2014 (31 Dec. 2014), pp.1-9, 9 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2014-11-02
Country of Publication
Egypt
No. of Pages
9
Main Subjects
Abstract EN
Conventional methods to determine the efficacy of bacteriophage (phage) for biocontrol of E.
coli require several days, due to the need to culture bacteria.
Furthermore, cell surface-attached phage particles may lyse bacterial cells during experiments, leading to an overestimation of phage activity.
DNA-based real-time quantitative polymerase chain reaction (qPCR) is a fast, sensitive, and highly specific means of enumerating pathogens.
However, qPCR may underestimate phage activity due to its inability to distinguish viable from nonviable cells.
In this study, we evaluated the suitability of propidium monoazide (PMA), a microbial membrane-impermeable dye that inhibits amplification of extracellular DNA and DNA within dead or membrane-compromised cells as a means of using qPCR to identify only intact E.
coli cells that survive phage exposure.
Escherichia coli O157:H7 strain R508N and 4 phages (T5-like, T1-like, T4-like, and O1-like) were studied.
Results compared PMA-qPCR and direct plating and confirmed that PMA could successfully inhibit amplification of DNA from compromised/damaged cells E.
coli O157:H7.
Compared to PMA-qPCR, direct plating overestimated (P < 0.01) phage efficacy as cell surface-attached phage particles lysed E.
coli O157:H7 during the plating process.
Treatment of samples with PMA in combination with qPCR can therefore be considered beneficial when assessing the efficacy of bacteriophage for biocontrol of E.
coli O157:H7.
American Psychological Association (APA)
Liu, Hui& Niu, Yan D.& Li, Jinquan& Stanford, Kim& McAllister, Tim A.. 2014. Rapid and Accurate Detection of Bacteriophage Activity against Escherichia coli O157:H7 by Propidium Monoazide Real-Time PCR. BioMed Research International،Vol. 2014, no. 2014, pp.1-9.
https://search.emarefa.net/detail/BIM-1016222
Modern Language Association (MLA)
Liu, Hui…[et al.]. Rapid and Accurate Detection of Bacteriophage Activity against Escherichia coli O157:H7 by Propidium Monoazide Real-Time PCR. BioMed Research International No. 2014 (2014), pp.1-9.
https://search.emarefa.net/detail/BIM-1016222
American Medical Association (AMA)
Liu, Hui& Niu, Yan D.& Li, Jinquan& Stanford, Kim& McAllister, Tim A.. Rapid and Accurate Detection of Bacteriophage Activity against Escherichia coli O157:H7 by Propidium Monoazide Real-Time PCR. BioMed Research International. 2014. Vol. 2014, no. 2014, pp.1-9.
https://search.emarefa.net/detail/BIM-1016222
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-1016222