Non-competitive fluorescence polarization immunoassay for detection of H5 avian influenza virus using a portable analyzer

Abstract

Nowadays, the diagnosis of viral infections is receiving broad attention. We have developed a non-competitive fluorescence polarization immunoassay (NC-FPIA), which is a separation-free immunoassay, for a virus detection. H5 subtype avian influenza virus (H5-AIV) was used as a model virus for the proof of concept. The fluorescein-labeled Fab fragment that binds to H5 hemagglutinin was used for NC-FPIA. The purified H5-AIV which has H5 hemagglutinin was mixed with the fluorescein-labeled Fab fragment. After that, the degree of fluorescence polarization was measured with a portable FPIA analyzer. H5-AIV was successfully detected with an incubation time of 15 min. In addition, the portable FPIA analyzer enables performance of on-site NC-FPIA with a sample volume of 20 μL or less. This is the first research of detecting a virus particle by FPIA. This NC-FPIA can be applied to rapid on-site diagnosis of various viruses. Supplementary Information The online version contains supplementary material available at 10.1007/s00216-021-03193-y.

Non-competitive fluorescence polarization immunosensing for CD9 detection using a peptide as a tracer

“…However, the Δ P was only 18.7 m P for ×10, 16.7 m P for ×100, and 11.7 m P for ×1000 samples. A similar tendency for Δ P , which is much smaller than expected, has been observed in virus detection using antibody fragments such as Fab 22 and scFV 38 as tracers. For large targets such as particles and viruses, it seems that the theoretical calculation of P values must be modified or reconstructed.…”

“…[18][19][20][21] This method can also be applied to virus detection. 22 The NC-FPIA is even simpler than the conventional FPIA. If a tracer that is fluorescently labeled with a substance that has an affinity for the target can be produced, this method can be applied to the detection of various proteins, antibodies, viruses, and so forth.…”

“…The sensitivity of the NC-FPIA also depends on the change in the molecular weight of the tracer before and after binding to the target, as in the conventional FPIA. The tracers we have prepared so far are a H5 hemagglutinin fragment (∼30 kDa) for anti-H5 avian influenza antibody detection, 18 a VHH antibody (∼15 kDa) for rabbit IgG detection, 19 Fab fragments (∼50 kDa) for C-reactive protein (CRP) detection 20 and avian influenza virus (H5N3) detection, 22 and a spike protein fragment (∼39 kDa) for anti-SARS-CoV-2 antibody detection. 21 On the other hand, peptides have been widely studied as drugs for metabolic and tumor related diseases 23 and as valuable tools for diagnostic applications.…”

Development of a Rapid Fluorescent Diagnostic System for Early Detection of the Highly Pathogenic Avian Influenza H5 Clade 2.3.4.4 Viruses in Chicken Stool

“…Multiple research groups have been focused on the development of mAbs for detecting H5 influenza viruses, and the LoD of H5-specific RDTs currently ranges from 0.23 ng/mL to 2.8 μg/mL for HA antigen detection [ 19 , 46 ]. The present study could not confirm or validate the detection of H5 HPAIVs within the 2.3.4.4 clade using commercial RDTs.…”

Facile and Unplugged Surface Plasmon Resonance Biosensor with NIR-Emitting Perovskite Nanocomposites for Fast Detection of SARS-CoV-2

“…Although biosensors using advanced sensing technology allow for accurate detection of most pathogens, detection methods for highly pathogenic viruses have been primarily centralized in large labs due to the need for complex instrumentation and well-trained technicians. − Consequently, people have to travel to hospitals with large labs for diagnosis and often wait for several days for results . However, the global pandemic of coronavirus disease-2019 (COVID-19) in recent years tells us that an easy-to-operate, field-deployable biosensing platform could provide a great help in preventing the spread of the disease − since portable platforms could be transported to regions lacking large labs for on-site detection .…”