tetano
Editor, Senior Moderator
Appl Microbiol Biotechnol. 2016 Jul 2. [Epub ahead of print]
[h=1]Multiplex assay for subtyping avian influenza A viruses by cDNA hybridization and adapter-mediated amplification.[/h] Yang G[SUP]1[/SUP], Jones J[SUP]1[/SUP], Jang Y[SUP]1[/SUP], Davis CT[SUP]2[/SUP].
[h=3]Author information[/h]
[h=3]Abstract[/h] Multiple subtypes of influenza A viruses circulating in animals must be closely monitored to understand their risk to humans and animal populations. Many molecular-based subtyping methods require constant monitoring of viral genomes for primer and/or probe mismatches and are prone to primer-primer interactions. This report presents a new approach that involves target enrichment through cDNA hybridization followed by adapter-mediated amplification for subtyping influenza virus (AmASIV). As a proof of concept, the AmASIV assay was multiplexed to specifically detect and differentiate influenza A virus subtypes (H5, N5, N7, and N9) in a single reaction without cross-recognition of nontarget subtypes or influenza B virus. The limit of detection (LOD) of AmASIV, as measured by 50 % egg-infective dose per reaction (EID[SUB]50[/SUB]/reaction), was comparable to that of singleplex TaqMan? qPCR assays with LODs of 10[SUP]-0.6[/SUP] (H5), 10[SUP]2[/SUP] (N5), 10[SUP]-0.3[/SUP] (N7), and 10[SUP]-0.5[/SUP] (N9) EID[SUB]50[/SUB]/reaction. The AmASIV will strengthen animal influenza virus surveillance and laboratory capacity to improve prevention and control of influenza.
[h=4]KEYWORDS:[/h] AmASIV; Influenza; Subtyping; Surveillance
PMID: 27372075 DOI: 10.1007/s00253-016-7664-8
[PubMed - as supplied by publisher]
[h=1]Multiplex assay for subtyping avian influenza A viruses by cDNA hybridization and adapter-mediated amplification.[/h] Yang G[SUP]1[/SUP], Jones J[SUP]1[/SUP], Jang Y[SUP]1[/SUP], Davis CT[SUP]2[/SUP].
[h=3]Author information[/h]
[h=3]Abstract[/h] Multiple subtypes of influenza A viruses circulating in animals must be closely monitored to understand their risk to humans and animal populations. Many molecular-based subtyping methods require constant monitoring of viral genomes for primer and/or probe mismatches and are prone to primer-primer interactions. This report presents a new approach that involves target enrichment through cDNA hybridization followed by adapter-mediated amplification for subtyping influenza virus (AmASIV). As a proof of concept, the AmASIV assay was multiplexed to specifically detect and differentiate influenza A virus subtypes (H5, N5, N7, and N9) in a single reaction without cross-recognition of nontarget subtypes or influenza B virus. The limit of detection (LOD) of AmASIV, as measured by 50 % egg-infective dose per reaction (EID[SUB]50[/SUB]/reaction), was comparable to that of singleplex TaqMan? qPCR assays with LODs of 10[SUP]-0.6[/SUP] (H5), 10[SUP]2[/SUP] (N5), 10[SUP]-0.3[/SUP] (N7), and 10[SUP]-0.5[/SUP] (N9) EID[SUB]50[/SUB]/reaction. The AmASIV will strengthen animal influenza virus surveillance and laboratory capacity to improve prevention and control of influenza.
[h=4]KEYWORDS:[/h] AmASIV; Influenza; Subtyping; Surveillance
PMID: 27372075 DOI: 10.1007/s00253-016-7664-8
[PubMed - as supplied by publisher]