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Product Manual BactoReal® Kit American Foulbrood Download PDF
Material Safety Data Sheet BactoReal® Kit American Foulbrood Download PDF
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Background:  American foulbrood (AFB) is a bacterial brood disease of honeybees caused by the Gram-positive and spore-forming bacterium Paenibacillus larvae. The disease is transferred and initiated only by the spore stage of the bacterium. Young bee larvae are infected by spores, then the spores germinate in the gut of the larva and the vegetative form of the bacteria begins to grow, taking its nourishment from the larva. Infected larvae normally die after their cell is sealed. The vegetative form of the bacterium produces many millions of spores, which may remain dormant for many years.

For detection of the​ Paenibacillus larvae genotype ERIC I, ingenetix offers an individual real-time PCR kit.

Description:  BactoReal® Kit American Foulbrood is a real-time PCR for the qualitative detection of the 16S rRNA gene of Paenibacillus larvae. 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 American Foulbrood
Order Number Reactions Channel Pathogen Channel IPC Target
DVEB05413 100 FAM Cy5 16S rRNA
DVEB05453 50 FAM Cy5 16S rRNA
For veterinary use only
Shipping Temperature: -20 °C to +4 °C
Product Features:
  • Amplification and detection: 16S rRNA gene of Paenibacillus larvae
  • 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) or Assay-only

PCR platforms: BactoReal® Kit American Foulbrood 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)).

Assay: Optional - this product is also available as an assay-only format with primers, probe and a positive control, but without reaction mix.

BactoReal® American Foulbrood
Order Number Reactions Channel Pathogen Channel IPC Target
RTGV924 100 FAM Cy5 16S rRNA gene
For veterinary use only
Shipping Temperature: -20 °C to +4 °C
Frequently Asked Questions:

The kit amplifies the 16S rRNA gene, with pathogen DNA detected in the FAM channel and the IPC in the Cy5 channel.
Infection occurs when larvae ingest spores of Paenibacillus larvae. The spores germinate in the gut, and the bacteria proliferate, eventually killing the larva.
The IPC is added during DNA extraction or PCR setup and ensures that negative results are not caused by failed extraction or PCR inhibition.
Validated instruments include ABI 7500, LightCycler® 480 II, and Mx3005P®. The kit is also compatible with other platforms detecting FAM and Cy5.
Strict workflow separation, sterile consumables, and negative extraction controls are recommended to avoid DNA contamination.
BactoReal®, MycoReal®, ParoReal® and ViroReal® Kits run with the same thermal cycling conditions.