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PCR Test for the Detection of Lawsonia intracellularis


418020

Código: 418020. Categorías: , , .
   
Descripción

1. Purpose of the test
ADIAVET™ LAW REAL TIME kit is intended to detect Lawsonia intracellularis using real-time
Polymerase Chain Reaction (PCR) technology from swab, tissue and faeces specimens of pig.
2. Lawsonia intracellularis
Proliferative enteropathy (PE) also known as proliferative ileitis, caused by the obligate intracellular
bacterium Lawsonia intracellularis is a world widespread disease. It has been reported in a variety of
species including pigs, horses, rabbits, hamsters…
The disease has two clinical manifestations on pigs: an acute haemorrhagic form often called porcine
hemorrhagic enteropathy (PHE) and a more chronic proliferative form often referred as porcine
intestinal adenomastosis (PIA).
Cases of acute hemorrhagic enteropathy occur most commonly in young adults 6 to 20 months old, but
older animals can also be affected.
Since PE attacks the lower small intestine (ileum), it impairs digestion and can lead to clinical signs.
Loose stools or watery diarrhoea may result along with loss of appetite, weight losses, reduce feed
efficiency and anaemia (Euzeby, 1995).
Histologically, the affected tissues show marked proliferation of epithelial cells of the intestinal crypts.
The magnitude of the proliferation varies widely but in developed lesions the wall is visibly thickened
and the overall diameter increased.
Lawsonia intracellularis has not been yet cultivated in cell-free media (Lawson et al., 1995). Gross
necropsy and laboratory examinations are frequently applied to diagnose PE. PCR detection of
Lawsonia intracellularis on faecal material is the only test allowing the detection of bacteria on live pig.
The PCR test is also the more sensitive test to detect Lawsonia intracellularis on intestinal mucosa
(Jones et al., 1993; Mc Orist et al., 1994).
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™ LAW REAL TIME kit enables the simultaneous detection of:
– L. intracellularis (probe labelled in FAM),
– 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 (Qiagen and Macherey-Nagel). Other
purification kits can be used if they have been validated by the user.

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