text.skipToContent text.skipToNavigation

cador TKP PCR Reagent

For multiplex identification of T. equigenitalis, K. pneumoniae, and P. aeruginosa DNA

  • Simultaneous identification of T. equigenitalis, K. pneumoniae, and P. aeruginosa DNA
  • Internal Control to monitor PCR inhibition
  • A full license for PCR without additional costs
  • Reagents developed with Veterinary Laboratories Agency (VLA), UK
The cador TKP PCR Reagent is for the identification of Taylorella equigenitalis (T. equigenitalis), Klebsiella pneumoniae (K. pneumoniae), and Pseudomonas aeruginosa (P. aeruginosa) DNA using real-time PCR on Rotor-Gene Q Systems. All 3 animal pathogens are identified in one tube on the Rotor-Gene Q. An Internal Control is included in the same tube, to monitor successful PCR amplification.

For more information, please contact us.

Cat No./ID: 285115
cador TKP PCR Reagent (96)
For 96 reactions: Pathogen Master Mix, QuantiTect Nucleic Acid Dilution Buffer, TKP Primer/Probes, TKP Control DNA
For Research Use Only. Not intended for any animal or human therapeutic or diagnostic procedures. No claim or representation is intended to provide information for the diagnosis, prevention or treatment of a disease. Regulatory requirements vary by country; product may not be available in your geographic area.

Performance
The cador TKP PCR Reagent contains reagents and enzymes for the specific and efficient amplification of highly conserved regions of the T. equigenitalis, K. pneumoniae, and P. aeruginosa genomes.

In addition, the cador TKP PCR Reagent contains a heterologous amplification system to identify possible PCR inhibition. Inhibition is determined by measuring the fluorescence signal in the yellow channel via amplification of an Internal Control (IC), which does not influence the amplification of the analytical PCR for T. equigenitalis, K. pneumoniae, and P. aeruginosa. An external positive control (TKP Control DNA, containing the targeted T. equigenitalis, K. pneumoniae, and P. aeruginosa DNA) is also supplied.

Analytical sensitivity was evaluated by analysis of DNA dilution series for each bacterium. Each real-time PCR experiment was performed in triplicate. The average lowest dilution at which amplification was seen on 3 separate occasions was designated the limit of detection (see the table Limit of detection for amplicons).

Table 1. Limit of detection for amplicons
Animal pathogen Limit of detection (gene copies/PCR)
K. pneumoniae 2.36
P. aeruginosa 2.52
T. equigenitalis 2.46
Several other pathogens that are not intended to be identified using the assay may also be present in the animal sample material. To test for potential cross-reactivity, the cador TKP PCR Reagent was used for analysis of other bacterial species (see the table Analysis of potential cross-reactivity). To test for interference, mixtures of the intended-use pathogens and potentially interfering bacteria were tested. No cross-reactivity or interference was observed.

Table 2. Analysis of potential cross-reactivity
Culture collection Reference Culture collection Reference
Actinobacillus equuli Not detected Pseudomonas putida Not detected
Actinobacillus pleuropneumoniae Not detected Rhodococcus equi Not detected
Arcano pyogenes Not detected Salmonella chittagong Not detected
Citrobacter freundi Not detected Salmonella nottingham Not detected
Enterobacter aerogenes Not detected Salmonella poona Not detected
Enterobacter cloacae Not detected Shigella flexneri Not detected
Enterococcus faecalis Not detected Staphylococcus aureus Not detected
Enterococcus faecium Not detected Staphylococcus epidermidis Not detected
Escherichia coli Not detected Streptococcus dysgalactiae Not detected
Escherichia fergusonii Not detected Streptococcus dysgalactiae equisimilus Not detected
Hafnia alvei Not detected Streptococcus equi Not detected
Oligella urethralis Not detected Streptococcus equi zooepidemicus Not detected
Pasteurella multocida Not detected Taylorella asinigenitalis Not detected
Proteus mirabilis Not detected Yersinia enterocolitica Not detected
Proteus morganis Not detected Yersinia fredericksenii Not detected
Pseudomonas fluorescens Not detected Yersinia rohdei Not detected
Principle
The cador TKP PCR Reagent is for the identification of T. equigenitalis, K. pneumoniae, and P. aeruginosa DNA using real-time PCR on Rotor-Gene Q Systems. All 3 animal pathogens are identified in one tube on the Rotor-Gene Q. An Internal Control is included in the same tube, to monitor successful PCR amplification.


Procedure
Animal pathogen identification by PCR is based on the amplification of specific regions of the pathogen genome. In real-time PCR, the amplified product can be detected via fluorescent dyes linked to oligonucleotide probes that bind specifically to the amplified product. Monitoring the fluorescence intensities during the PCR run (i.e., in real time) allows the identification of the accumulating product without having to reopen the reaction tubes after the PCR run, thus eliminating possible cross contamination. The amplicons for the 3 pathogens are detected by measuring the green, orange, and crimson fluorescence on a Rotor-Gene Q real-time PCR System (see the table Fluorescence channels for the different bacteria).

Table 3. Fluorescence channels for the different bacteria
Pathogen/Internal Control Channel                                
Klebsiella pneumoniae Green
Pseudomonas aeruginosa Orange
Taylorella equigenitalis Crimson
Internal Control Yellow
Applications
Reliable pathogen DNA identification by multiplex real-time PCR.
Features
Specifications
Quantitative/qualitative null
Recommended sample prep null
RUO/CE/ASR null
Sample type null
Thermal cycler null
What detected null

You are not authorized to download the resource

fragment fix placeholder

Customers who bought these products also bought