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Simultaneous quantitative detection of viable Escherichia coli O157:H7, Cronobacter spp., and Salmonella spp. using sodium deoxycholate-propidium monoazide with multiplex real-time PCR
- Source :
- Journal of Dairy Science. 102:2954-2965
- Publication Year :
- 2019
- Publisher :
- American Dairy Science Association, 2019.
-
Abstract
- Escherichia coli O157:H7, Cronobacter spp., and Salmonella spp. are common food-borne pathogens in milk that may cause serious diseases. In the present study, we established a simple, rapid, and specific method to simultaneously detect viable E. coli O157:H7, Cronobacter spp., and Salmonella spp. in milk. Three specific genes, fliC from E. coli O157:H7, cgcA from Cronobacter spp., and invA from Salmonella spp., were selected and used to design primers and probes. False-positive results were eliminated with the use of a combined sodium deoxycholate (SD) and propidium monoazide (PMA) treatment. Using the optimized parameters, this SD-PMA treatment combined with multiplex real-time PCR (SD-PMA-mRT-PCR) detected E. coli O157:H7, Cronobacter spp. and Salmonella spp. respectively, at 102 cfu/mL in pure culture or artificially spiked skim milk samples. A reasonable recovery rate (from 100 to 107%) for detection of viable bacteria using the SD-PMA-mRT-PCR assay was obtained in the presence of dead bacteria at 107 cfu/mL. The SD-PMA-mRT-PCR method developed in this study can accurately detect and monitor combined contamination with E. coli O157:H7, Cronobacter spp., and Salmonella spp. in milk and milk products.
- Subjects :
- Azides
Salmonella
food.ingredient
Biology
Escherichia coli O157
Real-Time Polymerase Chain Reaction
medicine.disease_cause
Sensitivity and Specificity
Microbiology
03 medical and health sciences
fluids and secretions
food
Propidium monoazide
Skimmed milk
Genetics
medicine
Animals
Multiplex
Cronobacter
Escherichia coli
030304 developmental biology
0303 health sciences
0402 animal and dairy science
04 agricultural and veterinary sciences
Contamination
biology.organism_classification
040201 dairy & animal science
Milk
Food Microbiology
Animal Science and Zoology
Bacteria
Deoxycholic Acid
Propidium
Food Science
Subjects
Details
- ISSN :
- 00220302
- Volume :
- 102
- Database :
- OpenAIRE
- Journal :
- Journal of Dairy Science
- Accession number :
- edsair.doi.dedup.....32f848ea66c7f21f622b27f70fc319cc