tetano
Editor, Senior Moderator
Sci Rep. 2017 Mar 21;7:44924. doi: 10.1038/srep44924.
[h=1]Multiplex Reverse-Transcription Loop-Mediated Isothermal Amplification Coupled with Cascade Invasive Reaction and Nanoparticle Hybridization for Subtyping of Influenza A Virus.[/h] Chi Y[SUP]1,[/SUP][SUP]2[/SUP], Ge Y[SUP]2[/SUP], Zhao K[SUP]2[/SUP], Zou B[SUP]3[/SUP], Liu B[SUP]4[/SUP], Qi X[SUP]2[/SUP], Bian Q[SUP]2[/SUP], Shi Z[SUP]2[/SUP], Zhu F[SUP]2[/SUP], Zhou M[SUP]2[/SUP], Cui L[SUP]2[/SUP], Su C[SUP]1[/SUP].
[h=3]Author information[/h]
[h=3]Abstract[/h] Considering the fatal human victims and economic loss caused by influenza virus infection every year, methodologies for rapid and on-site detection of influenza viruses are urgently needed. LAMP is the most commonly used nucleic acid isothermal amplification technology suitable for on-site use. However, for multiplex LAMP, differentiation of the amplicons derived from multiple targets is still challengeable currently. Here we developed a multiplex RT-LAMP assay for simultaneous amplification of three prominent subtypes of influenza viruses (A/H5, A/H7 and 2009A/H1). The amplicons were further identified by cascade invasive reaction and nanoparticle hybridization in separate target-specific detection tubes (referred to as mRT-LAMP-IRNH). The analytic sensitivities of the assay are 10 copies of RNA for all the three HA subtypes, and the specificity reached 100%. Clinical specimen analysis showed this assay had a combined sensitivity and specificity of 98.1% and 100%, respectively. Overall, the mRT-LAMP-IRNH assay can be used as a cost-saving method that utilizes a simple instrument to detect A/H5, A/H7, and 2009A/H1 influenza viruses, especially in resource-limited settings.
PMID: 28322309 PMCID: PMC5359610 DOI: 10.1038/srep44924
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[h=1]Multiplex Reverse-Transcription Loop-Mediated Isothermal Amplification Coupled with Cascade Invasive Reaction and Nanoparticle Hybridization for Subtyping of Influenza A Virus.[/h] Chi Y[SUP]1,[/SUP][SUP]2[/SUP], Ge Y[SUP]2[/SUP], Zhao K[SUP]2[/SUP], Zou B[SUP]3[/SUP], Liu B[SUP]4[/SUP], Qi X[SUP]2[/SUP], Bian Q[SUP]2[/SUP], Shi Z[SUP]2[/SUP], Zhu F[SUP]2[/SUP], Zhou M[SUP]2[/SUP], Cui L[SUP]2[/SUP], Su C[SUP]1[/SUP].
[h=3]Author information[/h]
[h=3]Abstract[/h] Considering the fatal human victims and economic loss caused by influenza virus infection every year, methodologies for rapid and on-site detection of influenza viruses are urgently needed. LAMP is the most commonly used nucleic acid isothermal amplification technology suitable for on-site use. However, for multiplex LAMP, differentiation of the amplicons derived from multiple targets is still challengeable currently. Here we developed a multiplex RT-LAMP assay for simultaneous amplification of three prominent subtypes of influenza viruses (A/H5, A/H7 and 2009A/H1). The amplicons were further identified by cascade invasive reaction and nanoparticle hybridization in separate target-specific detection tubes (referred to as mRT-LAMP-IRNH). The analytic sensitivities of the assay are 10 copies of RNA for all the three HA subtypes, and the specificity reached 100%. Clinical specimen analysis showed this assay had a combined sensitivity and specificity of 98.1% and 100%, respectively. Overall, the mRT-LAMP-IRNH assay can be used as a cost-saving method that utilizes a simple instrument to detect A/H5, A/H7, and 2009A/H1 influenza viruses, especially in resource-limited settings.
PMID: 28322309 PMCID: PMC5359610 DOI: 10.1038/srep44924
Free full text