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PCR Test for the Detection of Contagious Equine Metritis Klebsiella and/or Pseudomonas


ADI511-100

Código: ADI511-100. Categorías: , , .
   
Descripción

1. Purpose of the test
ADIAVET™ CEMO KLEB/PSEUDO REAL TIME kit is intended to detect Klebsiella pneumoniae and
Pseudomonas aeruginosa using real-time Polymerase Chain Reaction (PCR) technology from swab and
semen specimen of equine and bacterial culture.
2. K. pneumoniae and P. aeruginosa
Contagious equine metritis (CEM) is an inflammatory disease of the proximal and distal reproductive
tract of the mare mainly caused by Taylorella equigenitalis . Others pathogens encountered in metritis,
Taylorella asinigenitalis, K. pneumoniae and P. aeruginosa are found (OIE 2012).
K. pneumoniae and P. aeruginosa are Gram-negative bacillus classified in the Enterobacteriaceae
family.
K. pneumoniae is non-motile and facultative anaerobic. P. aeruginosa is motile and aerobic.
They are ubiquitous bacteria found in human and in various animal species as well as in aquatic habitats
and in the soil.
CEM diagnostic is standardized (e.g. OIE Terrestrial manual 2012 Chapter 2.5.2 and U47-108). Whereas
K. pneumoniae and P. aeruginosa are not OIE notified bacteria, the standard sampling process is often
apply for their detection. Briefly, swabbing of genital tractus of mare and stallion is performed. Swab is
sending to laboratory for bacterial diagnostic. Bacterial isolation is performed onto selective agar
media. Serology method can be alternative way but interpretation is often delicate, becoming
increasingly obsolete outside epidemiological context.
A variety of discriminatory real time PCR assays for rapid, sensitive and specific detection for K.
pneumoniae and P. aeruginosa directly from genital swabs without a need for additional bacterial
isolation have been developed. PCR addresses most of the associated short-comings of bacteriology
with fewer false negatives due to enhanced sensitivity and discriminatory specificity in detecting
bacterial presence despite factors including lower bacterial concentrations and heavily contaminated,
damaged or delayed samples.
This PCR can be realized at the same time with the ADIAVET™ CEMO TAYLORELLA REAL TIME PCR kit
for the detection of T. equigenitalis and T. asinigenitalis.
3. 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™ CEMO KLEB/PSEUDO REAL TIME kit enables the simultaneous detection of:
– K. pneumoniae (FAM labeled probe)
– P. aeruginosa (Cy5 labeled probe)
– an internal control of amplification step specific from an exogenous DNA (probe labelled
with a fluorochrome read in the same spectra as VIC and HEX).
Adiagene recommends using this test with DNA purification kits (Adiagene, Qiagen and Macherey-
Nagel). Other purification kits can be used if they have been validated by the user.

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