Quantification of Salmonella in Raw Poultry Using Droplet Digital PCR With a Whole Cell, Enrichment-free Approach
Salmonella contamination in poultry remains a persistent public health challenge, despite ongoing regulatory efforts to reduce its presence. Traditional culture-based enrichment methods commonly employed for detecting Salmonella have inherent limitations, such as biases in bacterial growth and inacc...
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| Language: | English |
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Elsevier
2025-04-01
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| Series: | Journal of Food Protection |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S0362028X2500050X |
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| author | Cheryl M. Armstrong Chin-Yi Chen Yicheng Xie Javier Atencia Sophie Pierre Yiping He Joe Lee Gretchen E. Dykes Kathleen Johnson Brice Froment Sean Martinos Joseph Capobianco |
| author_facet | Cheryl M. Armstrong Chin-Yi Chen Yicheng Xie Javier Atencia Sophie Pierre Yiping He Joe Lee Gretchen E. Dykes Kathleen Johnson Brice Froment Sean Martinos Joseph Capobianco |
| author_sort | Cheryl M. Armstrong |
| collection | DOAJ |
| description | Salmonella contamination in poultry remains a persistent public health challenge, despite ongoing regulatory efforts to reduce its presence. Traditional culture-based enrichment methods commonly employed for detecting Salmonella have inherent limitations, such as biases in bacterial growth and inaccurate quantification of initial contamination levels. This study introduces an enrichment-free approach for quantifying Salmonella in raw poultry, utilizing the Pathotrak system for bacterial separation and concentration, followed by Droplet Digital PCR (ddPCR) for absolute quantification. Our findings highlight that whole-cell preparations within ddPCR droplets yield significantly higher detection efficiency compared to extracted DNA, likely due to the reduced loss of genetic material during processing. The method was validated by correlating theoretical inoculation levels with ddPCR measurements, resulting in a strong linear relationship (R2 = 0.903) and a slope of 0.99, indicating high accuracy. Residuals from the regression model were normally distributed, confirming its validity. This platform offers a robust, rapid, and precise tool for detecting low levels of Salmonella in poultry products without enrichment, making it a more accurate and efficient approach to bacterial quantification in complex food matrices. |
| format | Article |
| id | doaj-art-a0f840edd88e48bcb9e6cdb4c28000c8 |
| institution | OA Journals |
| issn | 0362-028X |
| language | English |
| publishDate | 2025-04-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Journal of Food Protection |
| spelling | doaj-art-a0f840edd88e48bcb9e6cdb4c28000c82025-08-20T02:27:11ZengElsevierJournal of Food Protection0362-028X2025-04-0188510049810.1016/j.jfp.2025.100498Quantification of Salmonella in Raw Poultry Using Droplet Digital PCR With a Whole Cell, Enrichment-free ApproachCheryl M. Armstrong0Chin-Yi Chen1Yicheng Xie2Javier Atencia3Sophie Pierre4Yiping He5Joe Lee6Gretchen E. Dykes7Kathleen Johnson8Brice Froment9Sean Martinos10Joseph Capobianco11Characterization and Interventions for Foodborne Pathogens, Agricultural Research Service, United States Department of Agriculture, Wyndmoor, Pennsylvania, United StatesCharacterization and Interventions for Foodborne Pathogens, Agricultural Research Service, United States Department of Agriculture, Wyndmoor, Pennsylvania, United StatesBio-Rad Laboratories, Hercules, California, United StatesPathotrak Inc., Laurel, Maryland, United StatesBio-Rad Laboratories, Marnes-la-Coquette, France; Corresponding authors.Characterization and Interventions for Foodborne Pathogens, Agricultural Research Service, United States Department of Agriculture, Wyndmoor, Pennsylvania, United StatesCharacterization and Interventions for Foodborne Pathogens, Agricultural Research Service, United States Department of Agriculture, Wyndmoor, Pennsylvania, United StatesCharacterization and Interventions for Foodborne Pathogens, Agricultural Research Service, United States Department of Agriculture, Wyndmoor, Pennsylvania, United StatesCharacterization and Interventions for Foodborne Pathogens, Agricultural Research Service, United States Department of Agriculture, Wyndmoor, Pennsylvania, United StatesBio-Rad Laboratories, Marnes-la-Coquette, FrancePathotrak Inc., Laurel, Maryland, United StatesCharacterization and Interventions for Foodborne Pathogens, Agricultural Research Service, United States Department of Agriculture, Wyndmoor, Pennsylvania, United States; Corresponding authors.Salmonella contamination in poultry remains a persistent public health challenge, despite ongoing regulatory efforts to reduce its presence. Traditional culture-based enrichment methods commonly employed for detecting Salmonella have inherent limitations, such as biases in bacterial growth and inaccurate quantification of initial contamination levels. This study introduces an enrichment-free approach for quantifying Salmonella in raw poultry, utilizing the Pathotrak system for bacterial separation and concentration, followed by Droplet Digital PCR (ddPCR) for absolute quantification. Our findings highlight that whole-cell preparations within ddPCR droplets yield significantly higher detection efficiency compared to extracted DNA, likely due to the reduced loss of genetic material during processing. The method was validated by correlating theoretical inoculation levels with ddPCR measurements, resulting in a strong linear relationship (R2 = 0.903) and a slope of 0.99, indicating high accuracy. Residuals from the regression model were normally distributed, confirming its validity. This platform offers a robust, rapid, and precise tool for detecting low levels of Salmonella in poultry products without enrichment, making it a more accurate and efficient approach to bacterial quantification in complex food matrices.http://www.sciencedirect.com/science/article/pii/S0362028X2500050XDroplet digital PCR (ddPCR)PoultryQuantificationSalmonellaSample Preparation |
| spellingShingle | Cheryl M. Armstrong Chin-Yi Chen Yicheng Xie Javier Atencia Sophie Pierre Yiping He Joe Lee Gretchen E. Dykes Kathleen Johnson Brice Froment Sean Martinos Joseph Capobianco Quantification of Salmonella in Raw Poultry Using Droplet Digital PCR With a Whole Cell, Enrichment-free Approach Journal of Food Protection Droplet digital PCR (ddPCR) Poultry Quantification Salmonella Sample Preparation |
| title | Quantification of Salmonella in Raw Poultry Using Droplet Digital PCR With a Whole Cell, Enrichment-free Approach |
| title_full | Quantification of Salmonella in Raw Poultry Using Droplet Digital PCR With a Whole Cell, Enrichment-free Approach |
| title_fullStr | Quantification of Salmonella in Raw Poultry Using Droplet Digital PCR With a Whole Cell, Enrichment-free Approach |
| title_full_unstemmed | Quantification of Salmonella in Raw Poultry Using Droplet Digital PCR With a Whole Cell, Enrichment-free Approach |
| title_short | Quantification of Salmonella in Raw Poultry Using Droplet Digital PCR With a Whole Cell, Enrichment-free Approach |
| title_sort | quantification of salmonella in raw poultry using droplet digital pcr with a whole cell enrichment free approach |
| topic | Droplet digital PCR (ddPCR) Poultry Quantification Salmonella Sample Preparation |
| url | http://www.sciencedirect.com/science/article/pii/S0362028X2500050X |
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