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

Ready-to-use system for the isolation and the detection of V. cholerae using end-point PCR without enrichment

  • Rapid isolation of high quality DNA from stool
  • 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. Surveillance of drug resistant pathogens
    2. Epidemiologial studies
    3. Field surveillance of pathogens
    4. Surveys

Items Cat. # Size
Vibrio cholerae PCR Detection Kit 38500 24 preps

Vibrio cholerae PCR Detection Kit

Vibrio cholerae is a comma-shaped, gram-negative bacterium. It is the cause of cholera in humans, which affects the upper small intestine. Transmission of the disease is mainly through contaminated food or water. Human subjects affected by cholera exhibit severe watery diarrhea and vomiting, caused by the cholera toxin produced by the bacterium. Many cases of cholera are life-threatening, as diarrhea and associated vomiting can lead to rapid dehydration and electrolyte loss. Even with the extensive research on its epidemiology, cholera still affects over 5 million people per year worldwide.

Norgen’s Vibrio cholerae PCR Detection Kit  is a ready-to-use system for the isolation and detection of Vibrio cholerae without enrichment.  First, the kit contains components for the rapid isolation of total DNA, including bacterial DNA, from stool samples using spin-column chromatography based on Norgen’s proprietary resin (Nucleic Acid Isolation).  Second,  the kit contains Vibrio cholerae 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 Vibrio cholerae Master Mix contains reagents and enzymes for the specific amplification of a 333 bp region of the Vibrio cholerae genome. In addition, Norgen’s Vibrio cholerae 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 Vibrio cholerae 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 V. cholerae 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 V. cholerea PCR Mastermix can detect ≤100 copies of isolated DNA
  • Purchase Primer Set and Controls Separately - The V. cholerae 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 Vibrio cholerae PCR Detection Kit was determined by analysing a dilution series of a V. cholerae 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 Vibrio cholerae PCR Detection Kit on 1X TAE 1.7% Agarose gel.
  • The linear range of Norgen’s Vibrio cholerae PCR Detection Kit has been determined to cover concentrations from 1 x 102 cfu/µL to at least 1 x 106 cfu/µL of isolated DNA

Norgen's Vibrio cholerae PCR Detection Kit Contents
Lysis Solution
Lysis Additive
Binding Solution
Wash Solution I
Wash Solution II
Elution Buffer
Bead Tubes
Mini Spin Columns
Collection Tubes
Elution tubes (1.7 mL)
V. cholerae 2x PCR Master Mix
Control 2x PCR Master Mix
Isolation Control (IsoC)
V. cholerae  Positive Control (PosC)
Nuclease Free-Water
Norgen’s DNA Marker
Product Insert

Vibrio cholerae PCR Primer Set and Controls Contents
V. cholerae Primer Set
V. cholerae 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 V. cholerae 2x PCR Master Mix, Control 2x PCR Master Mix, Isolation Control (IsoC), and V. cholerae 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.