Multiplex Reverse-Transcription Loop-Mediated Isothermal Amplification Coupled with Cascade Invasive Reaction and Nanoparticle Hybridization for Subtyping of Influenza A Virus

• Published in: Scientific reports 2017-03, Vol.7 (1), p.44924-44924, Article 44924
• Main Authors: Chi, Ying, Ge, Yiyue, Zhao, Kangchen, Zou, Bingjie, Liu, Bin, Qi, Xian, Bian, Qian, Shi, Zhiyang, Zhu, Fengcai, Zhou, Minghao, Cui, Lunbiao, Su, Chuan
• Format: Article
• Language: English
• Subjects: 631/326/2521; 692/4017; Humanities and Social Sciences; Hybridization; Influenza; Influenza A; multidisciplinary; Nanoparticles; Nucleic acids; Pandemics; Ribonucleic acid; RNA; Science; Viruses
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/srep44924

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.