
Technical data sheet (Coxiella burnetii - Real time)
Technical details
for ready to use real time PCR kit for Coxiella burnetii (100 Reactions)
Coxiella burnetii is the causative agent of zoonostic disease called
Q-fever. It is an obligate intracellular rickettsial micro organisms. It has
been found throughout the world except New Zealand. it comes in human beings,
farm animals (like cattle, sheep, goat), pet animals like dog, cat, wild
animals as well as athropods. Infected animals may not show the symptoms.
Therefore it is essential to identify such carriers. In USA, there is very
high percentage of milk probes positive for Coxiella burnetii in cattle.
Quantitive assays should cover wide range of concentrations since the absolute
coxiella load as well as its time course are important diagnostic parameters
for indicating the prognosis as well as therapy. Coxiella burnetii has been
found in spleen, liver, aborted fetus, lungs, blood etc. Coxiella burnetii can
be detected through cell culture methods as well as serological methods like
FAT, ELISA etc, but these all methods have one or other following drawbacks:
low sensitivity, cross reactions or need a long time for detection, therefore
PCR is merging as godstandard as this is highly sensitive and finished within
a few hours. With real time PCR, one can get the results within 3-4 hours as
there is no post PCR stage. In case one can run the known standards with PCR,
it can give you the exact number of copies present in the sample. In case the
quantititive standard is not available, one can compare 2 samples as one see
the bacterial load as curve during real time assay. Real time curves can be
helpful during the therapy and predict the success of the treatment.
In case the load is going down, it means that the therapy is going to be
successful, otherwise one has to think about other alternatives.
target gene: highly conserved region of IS gene of coxiella burnetii
Samples: blood, serum, plasma, cell culture, milk, spleen, aborted fetus
tissue and other body fluids. Our test can be used to detect Coxiella burnetii
in ticks .
Keypoints : It has been checked for cross reactions for related and non
related micro organisms like Toxoplasma gondii, Leptospira, Mycoplasma,
Neospora caninum, Treponema pallidum, different Babesia, Mycoplasma, negative
tick, negative human DNA, negative bird DNA and many more. It has been diluted
for serial dilutions. It needs isolated DNA, which can be used through
commercial kits as well as home made methods.
Detection limit: minimum : 100 genomes per assay (detection Ct is lower 36 (this
should be taken as positive); 36-40 Ct should be taken as weak positive.)
Contents of the kit: Polymerase, buffer, PCR master mix, nucleotides, primers,
probes (with 6-carboxyfluoresceine and 6-carboxytetramethylrhodamine),
positive control, negative control, molecular water and manual.
Storage: -20 degree
Recommendations: always run the two samples per probe. Species specific
internal control can be used.
Only for in vitro use
for veterinary use
only for research use for human probes
to be used through technical person.