For research use only and NOT intended for In vitro-diagnostics.

A ready-to-use system for the isolation and the detection of E. coli 0157:H7 in food using end-point PCR

  • Rapid isolation of high quality DNA from food products
  • Contains two ready-to-use 2X PCR Master Mixes
  • High sensitivity and specificity
  • Includes an isolation control and a PCR control
  • Primer set and controls also available separately
  • Ideal for use in:
    1.  Food Surveillance and Monitoring
    2.  Surveys

 

Items Cat. # Size
E. coli O157:H7 PCR Detection Kit 41300 24 Preps

E. coli O157:H7 PCR Detection Kit

E. coli O157:H7 is a rod-shaped, gram negative bacterium. It is an enterohemorrhagic strain of the common E. coli bacterium and infection by the O157:H7 strain is commonly associated with hemorrhagic colitis. E. coli O157:H7 is recognized by its somatic (cell wall) antigen (O157) and its flagella antigen (H7). In addition, E. coli O157:H7 is known to produce Shiga-like toxins, which cause severe symptoms. While most patients can recover from the infection, up to 15% of the patients may develop hemolytic uremic syndrome, a type of kidney failure that could be fatal. Infection of E. coli O157:H7 usually results from consumption of poorly prepared food including undercooked meat (particularly ground beef), untreated water or raw unpasteurized milk.

Norgen’s E. coli O157:H7 PCR Detection Kit is a ready-to-use system for the isolation and detection of E. coli O157:H7. First, the kit contains components for the rapid isolation of total DNA, including bacterial DNA, from milk samples or enriched food samples using spin-column chromatography based on Norgen’s proprietary resin (Nucleic Acid Isolation).  Second,  the kit contains E. coli O157:H7 Master Mix and controls to allow for PCR Amplification, 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 E. coli O157:H7 Master Mix contains reagents and enzymes for the specific amplification of two Shiga-like toxin regions (348 bp and 587 bp) of the E. coli O157:H7 genome. In addition, Norgen’s E. coli O157:H7 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 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 E. coli O157:H7 PCR Primer Set and Controls are also available separately for end-point PCR detection.

  • 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 E. coli O157:H7 in under 3 hours after sample enrichment.
  • Process Different Sample Types - The kit can be used to detect E. coli O157:H7 from milk samples and enriched food samples
  • 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 milk
  • Purchase Primer Set and Controls Separately - The E. coli O157:H7 Primer Set, as well as the positive control and negative control can be purchased separately

Linear Range

  • The linear range (analytical measurement) of Norgen’s E. coli O157:H7 PCR Detection Kit was determined by analysing a dilution series of a E. coli O157:H7 quantification standard ranging from 1 x 107 cfu/µl to 1 x 10-1 cfu/µl.
  • Each dilution has been tested in replicates (n = 4) using Norgen’s E. coli O157:H7 PCR Detection Kit on 1X TAE 1.7% Agarose gel.
  • The linear range of Norgen’s E. coli O157:H7 PCR Detection Kit has been determined to cover concentrations from 1 x 102 cfu/µl to at least 1 x 106 cfu/µl
  • Under the conditions of the Norgen’s E. coli O157:H7 DNA Isolation procedure, Norgen’s E. coli O157:H7 PCR Detection Kit covers a linear range from 1,000 cfu/mL milk to at least 1 x 107 cfu/mL milk.

Norgen’s E. coli O157:H7 PCR Detection Kit (Meat Products) Contents
Digestion Buffer
Lysis Solution
Binding Solution
Wash Solution I
Wash Solution II
Elution Buffer
Proteinase K
Lysozyme
Mini Filter Spin Columns
Collection Tubes
Elution tubes (1.7 mL) 
E. coli O157:H7  2x PCR Master Mix
Control 2x PCR Master Mix
Isolation Control (IsoC)
E. coli O157:H7 Positive Control (PosC)
Nuclease Free-Water |
Norgen’s DNA Marker
Product Insert 

E. coli O157:H7 PCR Primer Set and Controls Contents
E. coli O157:H7 Primers Set
E. coli O157:H7 Positive Control (PosC)
Nuclease Free-Water |

Storage Conditions and Product Stability
All buffers should be kept tightly sealed and stored at room temperature (15-25oC). Buffers can be stored for up to 1 year without showing any reduction in performance. The Lysozyme should be stored at -20°C upon arrival, and the Digestion Buffer should be stored at -20°C after addition of the Lysozyme. The Proteinase K should be stored at -20°C upon arrival and after reconstitution. These reagents should remain stable for at least 1 year when stored at these conditions. The E. coli O157:H7 2x PCR Master Mix, 2x PCR Master Mix, E. coli O157:H7  Isolation Control (IsoC), and E. coli O157:H7 Positive Control (PosC) should be kept tightly sealed and stored at -20oC. These can be stored for up to 1 year without showing any reduction in performance. Repeated thawing and freezing (> 2 x) of these reagents should be avoided, as this may reduce the sensitivity. If the reagents are to be used only intermittently, they should be frozen in aliquots.