Journal article
Development of a one-run real-time PCR detection system for pathogens associated with porcine respiratory diseases
JOURNAL OF VETERINARY MEDICAL SCIENCE, v 82(2)
Feb 2020
PMID: 31866601
Featured in Collection : UN Sustainable Development Goals @ Drexel
Abstract
The etiology of Porcine respiratory disease complex is complicated by infections with multiple pathogens, and multiple infections increase the difficulty in identifying the causal pathogen. In this present study, we developed a detection system of microbes from porcine respiratory by using TaqMan real-time PCR (referred to as Dempo-PCR) to screen a broad range of pathogens associated with porcine respiratory diseases in a single run. We selected 17 porcine respiratory pathogens (Actinobacillus pleuropneumoniae, Boldetella bronchiseptica, Haemophilus parasuis, Pasteurella multocida, Pasteurella multocida toxin, Streptococcus suis, Mycoplasma hyopneumoniae, Mycoplasma hyorhinis, Mycoplasma hyosynovie, porcine circovirus 2, pseudorabies virus, porcine cytomegalovirus, swine influenza A virus, porcine reproductive and respiratory virus US strain, EU strain, porcine respiratory coronavirus and porcine hemagglutinating encephalomyelitis virus) as detection targets and designed novel specific primer-probe sets for seven of them. In sensitivity test by using standard curves from synthesized DNA, all primer-probe sets showed high sensitivity. However, porcine reproductive and respiratory virus is known to have a high frequency of genetic mutations, and the primer and probe sequences will need to be checked at a considerable frequency when performing Dempo-PCR from field samples. A total of 30 lung samples from swine showing respiratory symptoms on six farms were tested by the Dempo-PCR to validate the assay's clinical performance. As the results, 12 pathogens (5 virus and 7 bacteria) were detected and porcine reproductive and respiratory virus US strain, Mycoplasma hyorhinis, Haemophilus parasuis, and porcine cytomegalovirus were detected at high frequency. These results suggest that Dempo-PCR assay can be applied as a screening system with wide detection targets.
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Details
- Title
- Development of a one-run real-time PCR detection system for pathogens associated with porcine respiratory diseases
- Publication Details
- JOURNAL OF VETERINARY MEDICAL SCIENCE, v 82(2)
- Publisher
- JAPAN SOC VET SCI; TOKYO
- Number of pages
- 0
- Grant note
- This work was partially supported by The Ministry of Agriculture, Forestry and Fisheries of Japan, Award Number: the Research Project for Improving Food Safety and Animal Health of the Ministry of Agriculture, Forestry and Fisheries of Japan (716057223).
- Resource Type
- Journal article
- Language
- English
- Academic Unit
- Drexel University
- Web of Science ID
- WOS:000522825700020
- Scopus ID
- 2-s2.0-85080842332
- Other Identifier
- 991021860776704721
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- Collaboration types
- Domestic collaboration
- International collaboration
- Web of Science research areas
- Veterinary Sciences