Development of a Real-Time Multiplex PCR Assay with Propidium Monoazide Treatment for Simultaneous Detection of Live Salmonella, and Salmonella Enteritidis, S. Typhimurium, S. Pullorum, and S. Gallinarum, in Rinse Water of Chicken Carcasses
| Autor: | So Youn Youn, Suk Chan Jung, Min-Su Kang, Ok Mi Jeong, Byung Kook Choi |
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| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Serotype Salmonella animal diseases Salmonella enteritidis 030106 microbiology Biology medicine.disease_cause Applied Microbiology and Biotechnology Virology Analytical Chemistry Microbiology 03 medical and health sciences 030104 developmental biology Real-time polymerase chain reaction Propidium monoazide Multiplex polymerase chain reaction TaqMan medicine Safety Risk Reliability and Quality Safety Research Pathogen Food Science |
| Zdroj: | Food Analytical Methods. 10:1681-1689 |
| ISSN: | 1936-976X 1936-9751 |
| DOI: | 10.1007/s12161-016-0716-y |
| Popis: | Salmonella is a major food-borne pathogen in humans and a cause of local or systemic disease in animals. Therefore, rapid and reliable methods to detect these poultry-associated Salmonella serotypes are necessary for efficient control of Salmonella in poultry. The present study aimed to develop a real-time multiplex PCR (MqPCR) method to simultaneously detect and/or differentiate Salmonella sp. and poultry-associated serotypes, including Salmonella Enteritidis, Salmonella Typhimurium, Salmonella Pullorum, and Salmonella Gallinarum. A MqPCR method was designed using four specific primer pairs and probes for the detection of Salmonella sp., including S. Enteritidis, S. Typhimurium, S. Pullorum, and S. Gallinarum. Additionally, a novel TaqMan-based MqPCR method combined with propidium monoazide (PMA) treatment was developed for the simultaneous quantification of viable cells of Salmonella sp. and these four Salmonella serotypes in rinse water of chicken carcasses. The MqPCR assay specifically detected Salmonella sp., S. Enteritidis, S. Typhimurium, S. Pullorum, and S. Gallinarum, showing 100% sensitivity and 100% specificity. This optimized PMA-MqPCR assay could detect live Salmonella (100–106 CFU/reaction) without enrichment in live/dead cell mixtures from spiked rinse water of chicken carcasses. The procedure for detecting live Salmonella required less than 2 h to complete. This PMA TaqMan-based MqPCR technique facilitates accurate and rapid monitoring of contamination with viable Salmonella. Also, the assay enables simultaneous identification of S. Enteritidis, S. Typhimurium, S. Pullorum, and S. Gallinarum in rinse water of chicken carcasses. The assay developed in this study will be useful in diagnostic laboratories for improving Salmonella control in poultry and poultry products. |
| Databáze: | OpenAIRE |
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