Multiplex Real-Time Polymerase Chain Reaction for Simultaneous Quantification of Salmonella spp., Escherichia coli, and Staphylococcus aureus in Different Food Matrices: Advantages and Disadvantages
Joint Authors
Lopes, Amanda Teixeira Sampaio
Albuquerque, George Rêgo
Maciel, Bianca Mendes
Source
Issue
Vol. 2018, Issue 2018 (31 Dec. 2018), pp.1-12, 12 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2018-09-25
Country of Publication
Egypt
No. of Pages
12
Main Subjects
Abstract EN
Quantitative real-time polymerase chain reactions (qPCRs) of the most prevalent bacteria causing foodborne diseases worldwide, such as Salmonella spp., Escherichia coli, and Staphylococcus aureus, can be an important tool for quantitative microbial risk assessment, which requires numerical data to determine the level of contamination at a specific stage of food production.
However, most of qPCR assays described in the literature for these pathogens are qualitative; their objective is pathogen detection and not pathogen quantification.
Thus, the aim of our work was to develop a qPCR for the simultaneous quantification of Salmonella spp., E.
coli, and S.
aureus and to propose its use in the analysis of foods, as a tool for microbiological quality monitoring.
For this, a multiplex qPCR was standardized for the simultaneous quantification of specific fragments of target genes (ssf, phoA, and nuc) corresponding to each one of the mentioned bacteria.
The limit of detection of the technique was 13, 10, and 12 gene copies for ssf, phoA, and nuc, respectively; standard curves showed R2 > 0.99, with efficiencies ranging from 99 to 110%, and inter- and intraexperiment reproducibility presented a low coefficient of variation in all trials.
This methodology was applied in different food matrices (milk, ground beef, and oyster meat), and the results were compared with official microbiological culture methodology and with ready-to-use test.
Advantages and disadvantages of each methodology used in this study are pointed out.
We suggest that this multiplex qPCR can be used as a rapid screening technique for the analysis of food microbiological quality.
American Psychological Association (APA)
Lopes, Amanda Teixeira Sampaio& Albuquerque, George Rêgo& Maciel, Bianca Mendes. 2018. Multiplex Real-Time Polymerase Chain Reaction for Simultaneous Quantification of Salmonella spp., Escherichia coli, and Staphylococcus aureus in Different Food Matrices: Advantages and Disadvantages. BioMed Research International،Vol. 2018, no. 2018, pp.1-12.
https://search.emarefa.net/detail/BIM-1127702
Modern Language Association (MLA)
Lopes, Amanda Teixeira Sampaio…[et al.]. Multiplex Real-Time Polymerase Chain Reaction for Simultaneous Quantification of Salmonella spp., Escherichia coli, and Staphylococcus aureus in Different Food Matrices: Advantages and Disadvantages. BioMed Research International No. 2018 (2018), pp.1-12.
https://search.emarefa.net/detail/BIM-1127702
American Medical Association (AMA)
Lopes, Amanda Teixeira Sampaio& Albuquerque, George Rêgo& Maciel, Bianca Mendes. Multiplex Real-Time Polymerase Chain Reaction for Simultaneous Quantification of Salmonella spp., Escherichia coli, and Staphylococcus aureus in Different Food Matrices: Advantages and Disadvantages. BioMed Research International. 2018. Vol. 2018, no. 2018, pp.1-12.
https://search.emarefa.net/detail/BIM-1127702
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-1127702