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• W1998003003 abstract "• GeXP-multiplex PCR assay developed to detect nine avian respiratory pathogens in one reaction. • This assay is a rapid method with high sensitivity and specificity for identifying very important avian influenza subtypes (H5, H7, H9) and six respiratory pathogens. • GeXP-multiplex PCR assay can be adopted for molecular epidemiologic survey of avian respiratory pathogens. 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." @default.
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• W1998003003 date "2014-10-01" @default.
• W1998003003 modified "2023-09-27" @default.
• W1998003003 title "Simultaneous typing of nine avian respiratory pathogens using a novel GeXP analyzer-based multiplex PCR assay" @default.
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• W1998003003 doi "https://doi.org/10.1016/j.jviromet.2014.07.007" @default.
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