Evaluation of a novel micro/nanofluidic chip platform for the detection of influenza A and B virus in patients with influenza-like illness

Abstract

We introduced a novel micro/nanofluidic chip platform (MNCP), which is based on an isothermal nucleic acid amplification method. This study aimed to evaluate the MNCP method for influenza A and B viruses detecting and subtyping using throat swab samples from patients with influenza-like illness (ILI). A total of 266 throat swab samples from 266 non-repeated patients with ILI were tested for influenza A and B viruses using three methods, MNCP, a rapid influenza diagnostic test (RIDT), and real-time reverse transcription polymerase chain reaction (rRT-PCR). The results of MNCP were compared to those obtained by rRT-PCR and RIDT and the performance of MNCP was further evaluated. Compared with rRT-PCR results, the rates of sensitivity, specificity, overall concordance, and the kappa value of MNCP were 98.89%, 96.97%, 97.65%, and 0.95 for influenza A virus; 94.95%, 99.38%, 97.68%, and 0.95 for influenza B virus, respectively. Subtypes of influenza A viruses, e.g., A(H1N1)pdm09, A(H3N2), and A(not subtyped), and influenza B viruses could be distinguished in one MNCP assay within 1 h. Compared with rRT-PCR and MNCP, RIDT showed poor clinical sensitivity for influenza virus detection. This study showed MNCP is rapid, sensitive and versatile detecting system with potential for clinical application in pathogen diagnosis for patients with ILI.

Characterization of viral infections in children with influenza-like-illness during December 2018–January 2019

“…The method had been validated before it was incorporated in the study and could detect 18 common pathogens or subtypes within 45 minutes after loading the nucleic acid sample. Based on the isothermal amplification technology and the combined MNCP platform, the assay platform had been applied to detect ILI cases with influenza virus infection during the 2017-2018 seasonal influenza epidemic, with improved performance compared with a commercially available rRT-PCR test ( Li et al., 2019 ). After emergence of COVID-19, SARS-CoV-2 spike and nucleocapsid genes were incorporated into the system and the sequences of all the primers and probes can be found from published references ( Xing et al., 2020 ).…”

“…A high-throughput, multi-index isothermal amplification platform for rapid detection of 19 types of common respiratory viruses or subspecies, including SARS-CoV-2 had been reported in 2020, the assays based on nucleic acid sequence-based amplification (NASBA) and previously reported micro/nanofluidic chip platform (MNCP) ( Xing et al., 2020 ). Other multiple assay platforms based on real-time nucleic acid assay have been incorporated into routine diagnosis during and post the COVID-19 epidemic, broaden the understanding of multiple viral pathogenic infections ( Li et al., 2019 ).…”

The development of droplet-based microfluidic virus detection technology for human infectious diseases

“…Li et al introduced a novel microfluidic chip platform based on the isothermal nucleic acid amplification method. 16 Combining laser-induced fluorescence with microfluidic droplets is a significant advancement in the development of the digital reverse transcription loop-mediated isothermal amplification (RT-LAMP) platform for point-of-care droplet analysis. 17…”

“…However, vaccine development is a long-term process, and for emerging viral pathogens and viruses with high mutation frequency, vaccines obviously cannot achieve the effect of rapid disease control. 6 Therefore, rapid, accurate, and early detection of viral infections is a growing need for preventive medicine. Reverse transcription polymerase chain reaction (RT-PCR) is an amplification technique for specific DNA sequences and is a standard technique for viral diagnostics that has been used for decades.…”

Nanofluidic devices prepared by an atomic force microscopy-based single-scratch approach

“…Nanofluidic devices have drawn significant attention because of their potential application in various fields, such as health care, 1 medicine, 2 and DNA/protein transport. 3,4 Fabrication of nanofluidic chips using a simple and highly efficient approach has thus become a research focus and challenge for the application of nanofluidic devices.…”