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PCR Test for the Detection of Mycoplasma hyopneumoniae


418022

Código: 418022. Categorías: , , .
   
Descripción

1. Purpose of the test
ADIAVET™ M.HYOP REAL TIME kit is intended to detect Mycoplasma hyopneumoniae using real-time
Polymerase Chain Reaction (PCR) technology from tissue, tracheobronchiolar washing and oral fluid
specimens of pig, as well as from bacterial culture.
2. Pathogen
Mycoplasma hyopneumoniae is the primary agent of enzootic porcine pneumonia (EPP), one of the
most important direct or indirect causes of respiratory infectious diseases. The disease has a world wide
distribution and causes considerable economic losses in swine production due to reduce growth rate
and feed conversion efficiency. M. hyopneumoniae has a specific pathogenicity and leads to secondary
infection by other pathogenic bacteria such as Pasteurella multocida or Actinobacillus
pleuropneumoniae. The contamination occurs at each stage of piglet production, from sows to piglets
but also from pigs themselves by direct contact (Kobisch and Friis, 1996).
Diagnosis:
The isolation of M. hyopneumoniae by culture is not currently performed by diagnostic laboratories
because it is tedious and time consuming and may require as long as 1 month (Friis, 1975). Moreover, in
the respiratory tract of piglets it is often associated with bacteria (P. multocida, A. pleuropneumoniae)
as well as other mycoplasma species (M. hyorhinis and M. flocculare). Two methods are currently used
for EPP diagnosis: serological analysis such as blocking ELISA or detection of the organism on lung
sections by a direct immunofluorescent test using polyclonal antibodies (L’Ecuyer et Boulanger, 1970).
Many authors have described M. hyopneumoniae detection by PCR test. This technique allowed the
specific and rapid detection of mycoplasma directly on live pigs (Mattsson et al., 1995; Verdin et al .,
1996; Baumeister et al., 1998). The PCR test allowed the detection of M. hyopneumoniae on
seropositive pigs (infected or vaccinated pigs).
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™ M.HYOP REAL TIME kit enables the simultaneous detection of:
– Mycoplasma hyopneumoniae with probe labelled in FAM.
– The RNase P, 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).
ADIAGENE recommends using this test with DNA purification kits (Qiagen or Macherey-Nagel). Other
purification kits can be used if they have been validated by the user.

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