Evaluation of a Real-Time PCR Multiplex Assay For Detecting Foodborne Pathogenic Bacteria

İsmail Hakkı Tekiner; Haydar Özpınar

doi:10.17932/ IAU.IJFER.m.21495777.2015.1/1.7-16

Evaluation of a Real-Time PCR Multiplex Assay For Detecting Foodborne Pathogenic Bacteria

Abstract

Asbtract Molecular biological methods are feasible, quick and reliable tools of detecting major foodborne pathogens in food quality and safety monitoring. Among them, real-time PCR application is one of the testing platforms that optimally meets the criteria of performance against high throughput screening of microorganisms. The objective of this study was to evaluate the performance of a real-time PCR fourplex assay in simultaneous detection of major foodborne pathogenic bacteria, including Escherichia coli, Staphylococcus aureus, Listeria monocytogenes and Salmonella spp. To do this, the reference variants were inoculated to UHT milk sample individually as well as a cocktail containing them at all hands. All the spiked suspensions were initially exposed to pre-enrichment in buffered peptone water. Subsequently, 10-fold serial dilutions were prepared from them. The serially diluted suspensions were then transferred to Plate Count PC agar plates, followed by an aerobic incubation under the conditions according to the mannufacturer’s instructions. After that, the plates expressing a viable count of 1-10 CFU/ml, 10-100 CFU/ml and 100-1000 CFU/ ml were selected. The DNAs extracted by Eurofins GENESpin DNA isolation kit were subjected to a real-time PCR fourplex application using PowerChek™ Pathogen 4-plex detection kit Kogene Biotech, South Korea . The screening results revealed that the tested multiplex assay simultaneously exhibited its best performance as triplex for Sta, Lm and Sal in a concentration of 10-100 CFU/ml, excluding Ec. To conclude, detection and differentiation of multiplex bacteria are not yet optimized due to some technical limitations, and still evolving

Keywords

Food,Multiplex assay,real-time PCR,Sensitivity

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Details

Primary Language

English

Subjects

-

Journal Section

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Authors

İsmail Hakkı Tekiner This is me

Haydar Özpınar This is me

Publication Date

June 1, 2015

Submission Date

-

Acceptance Date

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Published in Issue

Year 2015 Volume: 1 Number: 1

DOI

https://doi.org/10.17932/ IAU.IJFER.m.21495777.2015.1/1.7-16

IZ

https://izlik.org/JA76PL43WD

Cite

RIS / Bibtex

APA

Tekiner, İ. H., & Özpınar, H. (2015). Evaluation of a Real-Time PCR Multiplex Assay For Detecting Foodborne Pathogenic Bacteria. International Journal of Food Engineering Research, 1(1), 7-16. https://doi.org/10.17932/ IAU.IJFER.m.21495777.2015.1/1.7-16

AMA

1.Tekiner İH, Özpınar H. Evaluation of a Real-Time PCR Multiplex Assay For Detecting Foodborne Pathogenic Bacteria. IJFER. 2015;1(1):7-16. doi:10.17932/ IAU.IJFER.m.21495777.2015.1/1.7-16

Chicago

Tekiner, İsmail Hakkı, and Haydar Özpınar. 2015. “Evaluation of a Real-Time PCR Multiplex Assay For Detecting Foodborne Pathogenic Bacteria”. International Journal of Food Engineering Research 1 (1): 7-16. https://doi.org/10.17932/ IAU.IJFER.m.21495777.2015.1/1.7-16.

EndNote

Tekiner İH, Özpınar H (June 1, 2015) Evaluation of a Real-Time PCR Multiplex Assay For Detecting Foodborne Pathogenic Bacteria. International Journal of Food Engineering Research 1 1 7–16.

IEEE

[1]İ. H. Tekiner and H. Özpınar, “Evaluation of a Real-Time PCR Multiplex Assay For Detecting Foodborne Pathogenic Bacteria”, IJFER, vol. 1, no. 1, pp. 7–16, June 2015, doi: 10.17932/ IAU.IJFER.m.21495777.2015.1/1.7-16.

ISNAD

Tekiner, İsmail Hakkı - Özpınar, Haydar. “Evaluation of a Real-Time PCR Multiplex Assay For Detecting Foodborne Pathogenic Bacteria”. International Journal of Food Engineering Research 1/1 (June 1, 2015): 7-16. https://doi.org/10.17932/ IAU.IJFER.m.21495777.2015.1/1.7-16.

JAMA

1.Tekiner İH, Özpınar H. Evaluation of a Real-Time PCR Multiplex Assay For Detecting Foodborne Pathogenic Bacteria. IJFER. 2015;1:7–16.

MLA

Tekiner, İsmail Hakkı, and Haydar Özpınar. “Evaluation of a Real-Time PCR Multiplex Assay For Detecting Foodborne Pathogenic Bacteria”. International Journal of Food Engineering Research, vol. 1, no. 1, June 2015, pp. 7-16, doi:10.17932/ IAU.IJFER.m.21495777.2015.1/1.7-16.

Vancouver

1.İsmail Hakkı Tekiner, Haydar Özpınar. Evaluation of a Real-Time PCR Multiplex Assay For Detecting Foodborne Pathogenic Bacteria. IJFER. 2015 Jun. 1;1(1):7-16. doi:10.17932/ IAU.IJFER.m.21495777.2015.1/1.7-16