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februarie
2015

A new PCR test for the detection of Actinobacillus pleuropneumoniae in the biological samples taken from swine is available at the Pasteur Romania Diagnosis Centre

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The code and name of the test:   PCR – App – Detection of Actinobacillus pleuropneumoniae by classical PCR

A new PCR test for the detection of Actinobacillus pleuropneumoniae in the biological samples taken from swine is available at the Pasteur Romania Diagnosis Centre.

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The Infectious Porcine Pleuropneumonia (IPP), caused by Actinobacillus pleuropneumoniae (App), can be found in pigs of all ages, including feral pigs, but the most common clinical symptoms are recorded in pigs older than 12 weeks during the finishing period. In the classical forms of the disease the animals exhibit fever and respiratory symptoms due to the characteristic lesions of necrotic haemorrhagic pneumonia or fibrinous pleuropneumonia. The disease may evolve supra-acutely, without clinical signs, with animals found dead, acutely, especially in immunologically naive flocks with losses by mortality of 15-20%, or sub-acutely with lower but variable mortality. In the case of endemically infected herds the disease becomes chronic in the surviving animals, affecting their welfare and causing a decrease in the zootechnical performance. Pigs can be carriers without developing any immune response. The disease generates significant economic losses.

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This PCR test is based on the specific amplification of the omlA gene encoding an outer membrane lipoprotein present in all the strains of Actinobacillus pleuropneumoniae, regardless of their serotype and apx pattern.

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The assay has been performed both on isolated reference strains of App in the field and on biological samples taken from swine.

art1-7The assay detection limit determined on the reference strains has proved to be 0.177 pg/ul DNA, as illustrated by the opposite figure.

Usefulness:

  • Confirming the causal agent of the disease (certainty diagnosis)
  • A tool in the analysis of the incidence of App in a herd of swine
  • A tool in preventing the spread of App within and between consignments of swine
  • An alternative diagnosis test in the case of negative bacteriological results due to recent treatments with antibiotics etc.
  • A tool in controlling the effectiveness of applied vaccines.

Bibliography:

Gram, T., Ahrens, P., 1998. Improved Diagnostic PCR Assay for Actinobacillus Pleuropneumoniae based on the Nucleotide Sequence of an Outer Membrane Lipoprotein, J. Clin. Microbiol. 36: 443–448.

Jessing Stine G., Angen Oystein, Inzana Tomas J. 2003, Evaluation of a Multiplex PCR Test for Simultaneous Identification and Serotyping of Actinobacillus Pleuropneumoniae Serotypes 2, 5, and 6, Journal of Clinical Microbiology 9 (41): 4095–4100

Angen Oystein, Ahrens Peter, Jessing Stine G. 2008, Development of a Multiplex PCR Test for Identification of Actinobacillus Pleuropneumoniae Serovars 1, 7, and 12, Veterinary Microbiology 132: 312–318

Specimens (refrigerated): Lungs, tonsils. Buffers for the lung, tonsils (impressions). Nasal swabs.

Response time: 2 business days

Methodology: quality test by classical PCR amplification

Reference values: N/A