Rapid Detection of Influenza Virus Subtypes Based on an Integrated Centrifugal Disc

Abstract

Influenza is a zoonotic disease, infecting a wide variety of warm-blooded animals. It is caused by an influenza virus, which has been found with hundreds of subtypes. These subtypes are often associated with different sources of infection and possess complex courses of infection. In the early stage of influenza infection, rapid subtype detection is very practicable to prevent the disease from getting worse. Herein, we presented a high-throughput microfluidic centrifugal disc for rapid detection of influenza virus subtypes. The disc realized detection reagent preloads, automated reagent control, and RT-LAMP detections. Six kinds of highly pathogenic influenza viruses could be simultaneously identified, including influenza A subtypes H1, H3, H5, H7, and H9 and influenza B virus. Two different fluorescent dyes could be used on the disc for real-time detection or read by the naked eye. The performance of the disc was demonstrated by testing the clinical samples. The integrated centrifugal disc was expected for rapid detection of influenza virus subtypes to facilitate accurate drug usage in resource-constrained settings and contribute to reduce the risk of the influenza pandemic.

Toward a next-generation diagnostic tool: A review on emerging isothermal nucleic acid amplification techniques for the detection of SARS-CoV-2 and other infectious viruses

“…Several reports have been published on microfluidic-based isothermal detection of infectious viruses [ 78 , [322] , [323] , [324] ], and most of these reports involve prototypes that are not fully developed for large-scale diagnosis. However, recently, a CRISPR/Cas-based preprint report named STOP (SHERLOCK Testing in One Pot) with potential POC advantages was published [ 275 ].…”

Advances and insights in the diagnosis of viral infections

“…Loop-mediated isothermal amplification method (LAMP) [ 235 ]—for DNR amplification at single incubation temperature using a set of four (two inners and two outer) unique primers to recognizing eight distinct sites of the target sequence. A DNA-displacing DNA polymerase (Bst or phi29 DNA polymerase) is used to initiate the synthesis, and the two primers form loop structures to facilitate and accelerate further amplification steps.…”

A Palm Germ-Radar (PaGeR) for rapid and simple COVID-19 detection by reverse transcription loop-mediated isothermal amplification (RT-LAMP)

“…Therefore, since its invention in 2000, a series of LAMP-based protocol had been successfully developed for detecting infectious diseases, such as avian influenza ( Imai et al, 2007 ), Middle East respiratory syndrome (MERS) ( Shirato et al, 2014 ), human immunodeficiency virus (HIV) ( Curtis et al, 2008 ) and African swine fever (ASF) ( Ye et al, 2019 ), etc. Some of them can achieve their applications in point-of-care testing (POCT) detection ( Nguyen et al, 2019 ; Yao et al, 2020 ).…”