BactoReal Kit Coxiella burnetii

Description

Cat# BC 243AQ

Manual
For use with the
 ABI PRISM® 7500 (Fast)
 Mx3005P®
 LightCycler® 480

 Product description
BactoReal® Kit Coxiella burnetii is a real-time PCR kit for detection of Coxiella burnetii DNA. This test was
developed for the ABI PRISM® 7500 (Fast) instrument (Thermo Fisher Scientific), LightCycler® 480 (Roche)
and for Mx3005P® (Agilent), but is also suitable for other real-time PCR instruments. This test allows the rapid
and sensitive detection of DNA of Coxiella burnetii from samples purified from tissues and swabs (e.g. with
the QIAamp DNA Mini Kit).
BactoReal® Kit Coxiella burnetii detects the IS1111 insertion sequence of Coxiella burnetii (target present at
least 20 times in the genome of Coxiella burnetii). A probe-specific amplification-curve at 530 nm (FAM
channel) indicates the amplification of Coxiella burnetii specific DNA.
An internal positive control system (IPC) for detection in VIC/HEX channel, (554 nm, order no. DVEB05811
or DVEB05851) or Cy5 channel (667 nm; order no. DVEB05813 or DVEB05853) excludes false-negative
interpretation of results due to inhibition of real-time PCR (see 8. Interpretation of PCR-data).
When using PCR-platforms not validated by ingenetix, an evaluation of the multiplex-PCR is recommended.
Please be aware that some PCR-platforms have to be calibrated with the corresponding dye before
performing multiplex-PCR.
BactoReal®
, MycoReal, ParoReal and ViroReal® Kits are optimized to run under the same thermal cycling
conditions. RNA and DNA material can be analysed in one run.

 Pathogen information
Coxiella burnetii (C. burnetii) is a Gram-negative, obligate intracellular, zoonotic bacterial pathogen, which is
the causative agent of Q fever (Coxiellosis). The disease is spread through inhalation of a spore-like cell
variant or close contact with body fluids or products from contaminated animals. Cattle, sheep and goats
represent the main animal reservoir. Seropositivity of ruminants for C. burnetti is associated with abortion,
stillbirth and fertility problems. The high resistance to standard disinfectants and environmental influences
renders disease control difficult. Antibiotic treatment and vaccination against Q fever is available.

3. Principle of real-time PCR
A specific DNA sequence of the pathogen genome is amplified and the generated PCR-product is detected
by an oligonucleotide-probe labelled with a fluorescent dye. This technology allows for a sequence-specific
detection of PCR amplificates.
 General Precautions
The user should always pay attention to the following:
 Always include a negative control per PCR-run (water instead of sample).
 Optional: for valid interpretation of results, a negative control should be included during DNA-extraction
(for example extraction of water instead of sample material), in order to exclude false-positive results due
to contamination with Coxiella burnetii DNA during extraction.
 Be careful when handling the positive control.
 Store and extract positive material (specimens, controls and amplicons) separately from all other reagents
and add it to the reaction mix in a spatially separated workspace.
 Periodically decontaminate benches and devices.
 Use sterile pipette tips with filters.
 Thaw all components thoroughly at room temperature before starting an assay. When thawed, mix the
components and centrifuge briefly.
 For MSDS, see www.ingenetix.com.