A Size-Selectively Biomolecule-Immobilized Nanoprobe-Based Chemiluminescent Lateral Flow Immunoassay for Detection of Avian-Origin Viruses

Abstract

In this study, a signal-amplifiable nanoprobe-based chemiluminescent lateral flow immunoassay (CL-LFA) was developed to detect avian influenza viruses (AIV) and other contagious and fatal viral avian-origin diseases worldwide. Signal-amplifiable nanoprobes are capable of size-selective immobilization of antibodies (binding receptors) and enzymes (signal transducers) on sensitive paper-based sensor platforms. Particle structure designs and conjugation pathways conducive for antigen accessibility to maximum amounts of immobilized enzymes and antibodies have advanced. The detection limit of the CL-LFA using the signal-amplifiable nanoprobe for the nucleoprotein of the H3N2 virus was 5 pM. Sensitivity tests for low pathogenicity avian influenza H9N2, H1N1, and high pathogenicity avian influenza H5N9 viruses were conducted, and the detection limits of CL-LFA were found to be 103.5 50% egg infective dose (EID50)/mL, 102.5 EID50/mL, and 104 EID50/mL, respectively, which is 20 to 100 times lower than that of a commercial AIV rapid test kit. Moreover, CL-LFA demonstrated high sensitivity and specificity against 37 clinical samples. The signal-amplifiable probe designed in this study is a potential diagnostic probe with ultrahigh sensitivity for applications in the field of clinical diagnosis, which requires sensitive antigen detection as evidenced by enhanced signaling capacity and sensitivity of the LFAs.

Avian Influenza in Wild Birds and Poultry: Dissemination Pathways, Monitoring Methods, and Virus Ecology

“…Immunofluorescent methods continue to improve and to evolve towards nanoscale. A micellar nanoparticle-based immunoluminescent lateral flow assay has been recently developed, able to detect a panel of avian influenza viruses such as H3N2, H9N2, H1N1, and H5N9 with detection limits of 10 3.5 EID50/mL, 10 2.5 EID50/mL, and 10 4 EID/mL, respectively [73].…”

Optical Biosensors for Diagnostics of Infectious Viral Disease: A Recent Update

“…To detect anti-DENV IgM antibodies, an indirect chemiluminescence enzyme immunoassay (CLEIA) based on synthetic peptides selected from the envelope protein of DENV using computational calculations was reported by Zhu et al [177]. For detection of avian influenza viruses (AIV) and other infectious and fatal viral avian-origin diseases, a signal-amplifiable nanoprobe-based chemiluminescent lateral flow immunoassay (CL-LFA) was developed by Zung et al [178]. In the approach, antibodies (binding receptors) and enzymes (signal transducers) can be immobilized on sensitive paper-based sensor platforms using size-selective signal amplifiable nanoprobes (Figure 4C).…”

Paper-based analytical devices for virus detection: Recent strategies for current and future pandemics

“…Another LFA based on a signal-amplifiable mesoporous silica nanoparticles (MSNs) was developed to detect avian influenza viruses (AIV) and other viral avian-origin diseases by Jung et al ( Fig. 2 A) [ 114 ]. Although conventional MSNs have been utilized owing to their properties such as porosity, high stability and biocompatibility, they have small pores (∼3 nm) which led to obtaining blocked surface during immobilization process due to the large size of biomolecules.…”

“…Reproduced with permission from Ref. [ 114 ] (B) (i) LFA for PRV gE-Ab detection. (ii) A smartphone with an ambient light sensor.…”

“… Target Fabrication Method Detection Method Label/Biorecognition element Linear Range LOD Ref. Hepatitis B virus (HBV) screen printing DPV Label free/acpcPNA 0.05-100 nM 1.45 pM [ 77 ] HIV type 1(HIV-1) p24 antigen wax printing Colorimetric anti-HIV-1 p24/anti-HIV-1 p24-HRP) Not specified 0.03 ng mL −1 [ 78 ] anti-SARV-CoV-2 IgG LFA Fluorescence lanthanide-doped polysterene nanoparticles (LNPs) Not specified N/A [ 113 ] avian influenza viruses (AIV), H9N2, H1N1, H5N9 LFA chemiluminescent PSNPs/antibody/HRP Not specified 10 3.5 10 2.5 10 4 EID 50 /mL [ 114 ] Porcine epidemic diarrhea virus (PEDV) LFA Fluorescence AuNPs/LFIS Not specified 0.055 μg mL -1 [ 40 ] Pseudorabies virus (PRV)-gE-Ab LFA Fluorescence Latex beads/ Not specified N/A [ 115 ] IL-6 Wax printing Colorimetric AuNPs-antibody 10 -3 -10 2 pg mL -1 10 -3 pg mL -1 in PBS, 1.3 pg mL -1 in blood [ 118 ] SARS-CoV-2 antibodies IgG and IgM Wax printing and screen printing SWV GO/spike protein receptor-binding domain (SP RBD) of SARS-CoV-2 1-10 3 ng mL -1 1 ng mL -...…”