<abstract xmlns="http://www.w3.org/1999/xhtml"><bold>Introduction:</bold> Porcine pleuropneumonia inflicts important economic losses on most commercial herds. Detection of subclinical or chronic infection in animals still remains a challenge, as isolation and identification of <italic>A. pleuropneumoniae</italic> serotypes is difficult and quantification of the bacteria on agar plates is often almost impossible. The aim of the study was to develop and evaluate a serotype-specific quantitative TaqMan probe-based PCR for detection of serotype 2 in pig lungs, tonsils, and nasal swabs.<bold>Material and Methods:</bold> The primers were designed from the capsular polysaccharide biosynthesis genes of <italic>A. pleuropneumoniae</italic> serotype 2. PCR specificity and sensitivity were evaluated using reference strains and several other bacterial species commonly isolated from pigs.<bold>Results:</bold> The real-time qPCR for detection of <italic>A. pleuropneumoniae</italic> serotype 2 was highly specific and gave no false positives with other serotypes or different bacterial species of pig origin. The detection limit for pure culture was 1.2 × 104 CFU/mL, for lung tissue and nasal swabs it was 1.2 × 105 CFU/mL, and for tonsils - 1.2 × 105 CFU/mL.<bold>Conclusion:</bold> The method can be used to serotype <italic>A. pleuropneumoniae</italic> isolates obtained during cultivation and to detect and identify <italic>A. pleuropneumoniae</italic> serotype 2 directly in nasal swabs and tonsil scrapings obtained from live pigs or lung tissue and tonsils collected post-mortem.</abstract>

Introduction: Porcine pleuropneumonia inflicts important economic losses on most commercial herds. Detection of subclinical or chronic infection in animals still remains a challenge, as isolation and identification of A. pleuropneumoniae serotypes is difficult and quantification of the bacteria on agar plates is often almost impossible. The aim of the study was to develop and evaluate a serotype-specific quantitative TaqMan probe-based PCR for detection of serotype 2 in pig lungs, tonsils, and nasal swabs.Material and Methods: The primers were designed from the capsular polysaccharide biosynthesis genes of A. pleuropneumoniae serotype 2. PCR specificity and sensitivity were evaluated using reference strains and several other bacterial species commonly isolated from pigs.Results: The real-time qPCR for detection of A. pleuropneumoniae serotype 2 was highly specific and gave no false positives with other serotypes or different bacterial species of pig origin. The detection limit for pure culture was 1.2 × 104 CFU/mL, for lung tissue and nasal swabs it was 1.2 × 105 CFU/mL, and for tonsils - 1.2 × 105 CFU/mL.Conclusion: The method can be used to serotype A. pleuropneumoniae isolates obtained during cultivation and to detect and identify A. pleuropneumoniae serotype 2 directly in nasal swabs and tonsil scrapings obtained from live pigs or lung tissue and tonsils collected post-mortem.

Real-time quantitative PCR for detection and identification of <italic>Actinobacillus pleuropneumoniae</italic> serotype 2

Department of Swine Diseases, National Veterinary Research Institute, 24-100 Pulawy,

Journal of Veterinary Research

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Real-time quantitative PCR for detection and identification of  serotype 2

Introduction: Porcine pleuropneumonia inflicts important economic losses on most commercial herds. Detection of subclinical or chronic infection in animals...

Real-time quantitative PCR for detection and identification of Actinobacillus pleuropneumoniae serotype 2

Published Online: Sep 23, 2016

Page range: 253 - 256

Received: Feb 02, 2016

Accepted: Jul 18, 2016

DOI: https://doi.org/10.1515/jvetres-2016-0038

Keywords
pigs, identification, quantitative real-time PCR, TaqMan probe

© 2016 Arkadiusz Dors et al., published by De Gruyter Open

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Real-time quantitative PCR for detection and identification of Actinobacillus pleuropneumoniae serotype 2

Published Online: Sep 23, 2016

Page range: 253 - 256

Received: Feb 02, 2016

Accepted: Jul 18, 2016

DOI: https://doi.org/10.1515/jvetres-2016-0038

Keywordspigs, identification, quantitative real-time PCR, TaqMan probe

© 2016 Arkadiusz Dors et al., published by De Gruyter Open

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Keywords
pigs, identification, quantitative real-time PCR, TaqMan probe