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A High-throughput Platform for the Screening of Salmonella spp./Shigella spp.
Published on: November 7, 2018
Development of a DNA macroarray for simultaneous detection of multiple foodborne pathogenic bacteria in fresh chicken
Chanida Kupradit1, Sureelak Rodtong, Mariena Ketudat-Cairns
1School of Biotechnology, Institute of Agricultural Technology, Suranaree University of Technology, Nakhon Ratchasima, 30000, Thailand.
Abstract:
A DNA macroarray was developed to provide the ability to detect multiple foodborne pathogens in fresh chicken meat. Probes targeted to the 16S rRNA and genus- and species-specific genes, including fimY, ipaH, prfA, and uspA, were selected for the specific detection of Salmonella spp., Shigella spp., Listeria monocytogenes, and Escherichia coli, respectively. The combination of target gene amplification by PCR and a DNA macroarray in our system was able to distinguish all target bacteria from pure cultures with a detection sensitivity of 10⁵ c.f.u. ml⁻¹. The DNA macroarray was also applied to 10 fresh chicken meat samples. The assay validation demonstrated that by combining the enrichment steps for the target bacteria and the DNA macroarray, all 4 target bacteria could be detected simultaneously from the fresh chicken samples. The sensitivity of L. monocytogenes and Shigella boydii detection in the fresh chicken samples was at least 10 and 3 c.f.u. of the initial contamination in 25 g samples, respectively. The advantages of our developed protocol are high accuracy and time reduction when compared to conventional culture. The macroarray developed in our investigation was cost effective compared to modern oligonucleotide microarray techniques because there was no expensive equipment required for the detection of multiple foodborne pathogens.
Insights
A novel DNA macroarray efficiently detects multiple foodborne pathogens like Salmonella and E. coli in chicken. This cost-effective method offers high accuracy and reduced time compared to traditional culture techniques.
Area of Science:
- Food Microbiology
- Molecular Diagnostics
- Food Safety
Background:
- Foodborne pathogens pose significant public health risks.
- Accurate and rapid detection methods are crucial for food safety, especially in poultry products.
- Conventional culture methods for pathogen detection are time-consuming and labor-intensive.
Purpose of the Study:
- To develop and validate a DNA macroarray system for simultaneous detection of multiple foodborne pathogens in fresh chicken meat.
- To assess the sensitivity and accuracy of the developed macroarray assay.
- To compare the efficiency of the macroarray method with conventional culture techniques.
Main Methods:
- Development of a DNA macroarray targeting 16S rRNA and specific genes (fimY, ipaH, prfA, uspA) for Salmonella spp., Shigella spp., Listeria monocytogenes, and Escherichia coli.
- Utilizing Polymerase Chain Reaction (PCR) for target gene amplification.
- Application of the DNA macroarray to pure bacterial cultures and spiked fresh chicken meat samples after enrichment.
Main Results:
- The PCR-amplified DNA macroarray system successfully distinguished all target bacteria from pure cultures with a sensitivity of 10⁵ c.f.u. ml⁻¹.
- Simultaneous detection of all four target pathogens was achieved in fresh chicken samples.
- The assay demonstrated high sensitivity for Listeria monocytogenes (≥10 c.f.u./25g) and Shigella boydii (≥3 c.f.u./25g).
Conclusions:
- The developed DNA macroarray assay provides a highly accurate and rapid method for detecting multiple foodborne pathogens in chicken meat.
- This cost-effective macroarray technique offers advantages over conventional culture methods, reducing analysis time and requiring no expensive equipment.
- The assay holds significant potential for enhancing food safety surveillance and control in the poultry industry.
