Salmonella enterica have emerged as significant foodborne pathogens that pose a serious public health problem. The symptoms of salmonellosis may include diarrhea, fever, vomiting, and abdominal cramps with elderly, new-born, and immunocompromised individuals the most susceptible. S. enterica is a facultatively anaerobic, Gram-negative bacterium that could survive low temperatures and freezing. It is a rare causative agent of mastitis in cow. However, Salmonella is commonly found in raw milk or dairy products made from unpasteurized milk. In addition, there is an increase in Salmonella contamination in milk after pasteurization, making milk an important vehicle of Salmonella transmission from infected farm animals to humans.
Norgen’s Milk Salmonella enterica PCR Detection Kit is a ready-to-use system for the isolation and detection of S. enterica. First, the kit contains components for the rapid isolation of total DNA, including bacterial DNA, from milk samples using spin-column chromatography based on Norgen’s proprietary resin (Nucleic Acid Isolation). Second, the kit contains S. enterica Master Mix and controls to allow for PCR Ampification, as well as a Control Master Mix to allow for amplification of both an Isolation Control and a PCR Control. The amplified PCR products are then detected using agarose gel electrophoresis. Alternatively, detection can be performed based on real-time PCR using melt curves (Detection). Please see the flowchart to the right.
The S. enterica Master Mix contains reagents and enzymes for the specific amplification of a 336 bp region of the S. enterica genome. In addition, Norgen’s Milk Salmonella enterica PCR Detection Kit contains a second Master Mix, the Control 2X PCR Master Mix, which can be used to identify possible PCR inhibition and/or inadequate isolation via a separate PCR reaction with the use of the provided PCR Control (PCRC) or Isolation Control (IsoC). This kit is designed to allow for the testing of 24 samples. The Salmonella enterica PCR Primer Set and Controls are also available separately for end-point PCR detection.
FEATURES AND BENEFITS
- Isolate High Quality, Inhibitor-Free DNA - Removal of highly concentrated salts, metabolic wastes and proteins provides high quality, concentrated DNA.
- Rapid Procedure - Isolate and detect S. enterica in under 3 hours.
- Accuracy in Interpreting Results – The amplification bands are clearly distinct, and the melt curves do not overlap
- High Sensitivity and Specificity - The limit of detection is 1000 cfu/mL of milk
- Purchase Primer Set and Controls Separately - The S. enterica Primer Set, as well as the positive control and negative control can be purchased separately