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PCR Test for the Detection of Q Fever


ADI143-100

Código: ADI143-100. Categorías: , , .
   
Descripción

1. Purpose of the test
ADIAVET™ COXIELLA REAL TIME kit is intended to detect and to quantify Coxiella burnetii using realtime
Polymerase Chain Reaction (PCR) technology from swab, tissue, faeces, amniotic fluid and milk
specimens of bovine, ovine and caprine.
2. Pathogen
Coxiella burnetii is a strictly intracellular Gram negative bacterium, agent of the Q fever (Burnet and
Freeman, 1937; Cox, 1938). The disease is present worldwide. C. burnetii is able to infect a large range
of hosts including humans. Q fever is a zoonose (pathology which can be transmitted to human) mainly
transmitted by ovine, bovine and caprine. Q fever usually results from inhalation of contaminated
aerosols originating mostly from dropping and body fluids of infected animals.
C. burnetii is present in reproductive apparatus, placenta and fluids produced during parturition or
abortion. The bacterium is excreted in milk and faeces of infected animals presenting no clinical signs.
Mostly infections are asymptomatic or subclinical by Human, but can be manifested as a flu-like
disease, or as hepatitis. A neurological involvement is also possible. Pericarditis and myocarditis are
rare. C. burnetii can induce abortions, causing economic losses in cattle. Prevention consists mostly in
precautions with pregnant females and with fluids from parturition.
Detection of C. burnetii by culture is long and difficult. Detection of antibodies doesn’t give
information on sanitary status of the animal.
The real-time PCR kit, ADIAVET™ COXIELLA REAL TIME, allows the identification of the bacteria C.
burnetii . It is applicable to a large number of samples and various biological matrixes.
3. Test performance
This PCR test was assessed against a panel of 130 issues from organisms preferencially found in the
same ecologic niche and/or close phylogeneticaly, among which Legionella. No crossing reaction was
observed.
The PCR detection limit is 1.5 C. burnetii /5μl PCR.
The PCR quantification limit is 2 C. burnetii /5μl PCR.
4. Description and purpose of the test
This test is based on enzymatic gene amplification or PCR technology.
Amplified products are detected in real-time thanks to a specific labelled hydrolysis probe (5’-
exonulease technology).
The ADIAVET™ COXIELLA REAL TIME kit enables the simultaneous detection of:
– The target Coxiella burnetii (probe labelled in FAM), specific of the IS1111 sequence
– GAPDH, an internal control of extraction and amplification steps specific from an
endogenous DNA (probe labelled with a fluorochrome read in the same spectra as VIC
and HEX).
An exogen control EPC-Ext added during the extraction of acellular samples allows validating
extraction and amplification steps (probe labelled with a fluorochrome read in the same spectra as VIC
and HEX).
The COX CTL+ contains a known amount of C. burnetii DNA. It allows the quantification of the positives
samples. The mesure unit is the equivalent genome (EG) number (or bacteria) per ml.
Each sample is analysed in a single well.
ADIAGENE validated the test using DNA purification kits (Qiagen, Macherey-Nagel). Other
purification kits can be used if they have been validated by the user.

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