tetano
Editor, Senior Moderator
J Virol Methods. 2014 Jul 12. pii: S0166-0934(14)00263-8. doi: 10.1016/j.jviromet.2014.07.007. [Epub ahead of print]
Simultaneous typing of nine avian respiratory pathogens using a novel GeXP analyzer-based multiplex PCR Assay.
Xie Z1, Luo S2, Xie L2, Liu J2, Pang Y2, Deng X2, Xie Z2, Fan Q2, Khan MI3.
Author information
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.
Copyright ? 2014. Published by Elsevier B.V.
KEYWORDS:
Avian respiratory pathogens; GenomeLab Gene Expression Profiler (GeXP); Multiplex PCR
PMID:
25025815
[PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/25025815
Simultaneous typing of nine avian respiratory pathogens using a novel GeXP analyzer-based multiplex PCR Assay.
Xie Z1, Luo S2, Xie L2, Liu J2, Pang Y2, Deng X2, Xie Z2, Fan Q2, Khan MI3.
Author information
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.
Copyright ? 2014. Published by Elsevier B.V.
KEYWORDS:
Avian respiratory pathogens; GenomeLab Gene Expression Profiler (GeXP); Multiplex PCR
PMID:
25025815
[PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/25025815