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Product Manual BactoReal® Kit Footrot Download PDF
Material Safety Data Sheet BactoReal® Kit Footrot Download PDF
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Background:  Footrot (infectious pododermatitis), is a hoof infection commonly found in sheep, goats, and cattle. It is responsible for the majority of cases of lameness in sheep. In sheep, there are two types of bacteria involved in causing footrot. Dichelobacter nodosus is the main cause of the disease, while Fusobacterium necrophorum is an associated pathogen. The two usually have a synergistic effect together. D. nodosus is an anaerobic, non-sporulating, Gram-negative bacterium. F. necrophorum, which is always present in the atmosphere, is an obligate anaerob, Gram-negative bacterium and produces a number of exotoxins that damage the hoof of the animal. Fusobacterium necrophorum is classified into subsp. necrophorum and subsp. funduliforme. Replication of the bacteria and disease transmission usually occur under humid and hot weather conditions that are typical of the rainy season.

​To differentiate between virulent and benign ​D. nodosus​ strains, ingenetix offers an additional real-time PCR test: ​BactoReal​®​ Typing Kit ​Dichelobacter nodosus.​

Description:  BactoReal® Kit Footrot is a real-time PCR for the qualitative detection of the pnpA gene of Dichelobacter nodosus and gyrB gene of Fusobacterium necrophorum. Our carefully designed primers and probe ensure highest sensitivity and specificity. The kit consists of an assay mix for the detection of the pathogen as well as a positive control and an internal positive control (IPC) system. To minimize PCR cross-contamination, the included reaction mix contains dUTP and uracil-N glycosylase (UNG).

BactoReal® Kit Footrot
Order Number Reactions Channel Pathogen Channel IPC Target
DVEB06813 100 FAM, VIC Cy5 pnpA/gyrB
DVEB06853 50 FAM, VIC Cy5 pnpA/gyrB
For veterinary use only
Shipping Temperature: -20 °C to +4 °C
Product Features:
  • Amplification and detection: pnpA gene of Dichelobacter nodosus and gyrB gene of Fusobacterium necrophorum.
  • Real-time PCR with rapid hot-start Taq DNA polymerase
  • ROX™ dye as passive reference
  • Internal Positive Control System to exclude false-negative results
  • Optimized to handle PCR inhibitors
  • PCR platforms: runs on all established standard real-time PCR platforms
  • Harmonized thermal profiles to run RNA and DNA samples simultaneously
  • Available formats: Kit (with reaction mix) and Assay (without reaction mix)

PCR platforms: BactoReal® Kit Footrot was validated with the ABI® 7500 Fast Real-Time PCR System (fast cycle parameters are not supported, Thermo Fisher Scientific), and was also tested with Mx3005P® (Agilent), qTOWER3G (Analytik Jena), MIC instrument (bio molecular systems), LightCycler® 480 II (Roche) and cobas Z 480 Analyzer (Roche). It is also compatible with other real-time PCR instruments which detect and differentiate fluorescence in the FAM and Cy5 channel (e.g. QuantStudio™ 5 / 6 / 7 (Thermo Fisher Scientific)).

BactoReal® Footrot
Order Number Reactions Channel Pathogen Channel IPC Target
RTGV941 100 FAM, VIC Cy5 pnpA/gyrB
Shipping Temperature: -20 °C to +4 °C
Frequently Asked Questions:

The kit detects DNA of Dichelobacter nodosus (pnpA gene) and Fusobacterium necrophorum subsp. necrophorum (gyrB gene) using multiplex real-time PCR.
D. nodosus is the primary cause of ovine footrot, while F. necrophorum contributes as a synergistic pathogen, increasing tissue damage and disease severity.
An artificial DNA internal positive control (IPC) is provided to monitor DNA extraction efficiency and detect possible PCR inhibition.
The kit is validated for DNA extracted from biopsies and swabs of affected hooves.
The detection limit is approximately 50 target DNA copies per PCR reaction, with validated specificity for D. nodosus and F. necrophorum subsp. necrophorum.
BactoReal®, MycoReal®, ParoReal® and ViroReal® Kits run with the same thermal cycling conditions.