Peer-reviewed veterinary case report
Simultaneous typing of nine avian respiratory pathogens using a novel GeXP analyzer-based multiplex PCR assay.
- Journal:
- Journal of virological methods
- Year:
- 2014
- Authors:
- Xie, Zhixun et al.
- Affiliation:
- Guangxi Veterinary Research Institute · China
- Species:
- bird
Plain-English summary
Researchers have developed a new test that can quickly identify nine different respiratory diseases in birds. This test can detect viruses like avian influenza and infectious bronchitis, as well as bacteria that cause respiratory issues. It was found to be very accurate when tested on 24 samples from birds with respiratory infections, showing better results than older testing methods. This new tool could help veterinarians diagnose and treat respiratory illnesses in birds more effectively. Overall, the new test works well for identifying these specific respiratory pathogens.
Abstract
A new, rapid, and high-throughput GenomeLab Gene Expression Profiler (GeXP) analyzer-based multiplex PCR method was developed for simultaneous detection and differentiation of nine avian respiratory pathogens. The respiratory pathogens included in this study were avian influenza subtypes H5, H7, and H9, infectious bronchitis virus (IBV), Newcastle disease virus (NDV), infectious laryngotracheitis virus (ILTV), Mycoplasma gallisepticum (MG), Mycoplasma synoviae (MS) and Haemophilus paragallinarum (HPG). Ten pairs of primers were designed using conserved and specific sequence genes of AIV subtypes and respiratory pathogens from GenBank. Single and mixed pathogen cDNA/DNA templates were used to evaluate the specificity of the GeXP-multiplex assay. The corresponding specific DNA products were amplified for each pathogen. The specific DNA product amplification peaks of nine respiratory pathogens were observed on the GeXP analyzer. Non-respiratory avian pathogens, including chicken infectious anemia virus, fowl adenovirus, avian reovirus and infectious bursal disease virus, did not produce DNA products. The detection limit for the GeXP-multiplex assay was determined to be 100 copies/μl using various pre-mixed plasmids/ssRNAs containing known target genes of the respiratory pathogens. Further, GeXP-multiplex PCR assay was 100% specific when 24 clinical samples with respiratory infections were tested in comparison with conventional PCR method. The GeXP-multiplex PCR assay provides a novel tool for simultaneous detection and differentiation of nine avian respiratory pathogens.
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Search related cases →Original publication: https://pubmed.ncbi.nlm.nih.gov/25025815/