tetano
Editor, Senior Moderator
Emerg Microbes Infect
. 2025 Dec 9:2602315.
doi: 10.1080/22221751.2025.2602315. Online ahead of print. Smartphone-assisted upconversion nanoparticle assay for rapid multiplex detection of H5, H7, and H10 avian influenza viruses
Jun Zhang[SUP] 1 [/SUP], Han Wu[SUP] 1 [/SUP], Ping Wang[SUP] 1 [/SUP], Jiamin Fu[SUP] 1 [/SUP], Xinyao Zheng[SUP] 2 [/SUP], Fang Wan[SUP] 3 [/SUP], Man Hu[SUP] 2 [/SUP], Fumin Liu[SUP] 1 [/SUP], Linfang Cheng[SUP] 1 [/SUP], Hangping Yao[SUP] 1 [/SUP], Nanping Wu[SUP] 1 [/SUP], Junli Zhu[SUP] 2 [/SUP], Haibo Wu[SUP] 1 [/SUP]
Affiliations
Avian influenza viruses (AIVs) of the H5, H7, and H10 subtypes pose substantial threats to global public health owing to their high pathogenicity, cross-species transmissibility, and potential to spark epidemics. Rapid and accurate detection is essential for outbreak control and zoonotic risk mitigation. Here, we report the development of a multiplex lateral flow immunoassay (LFA) based on core-shell upconversion nanoparticles (UCNPs) conjugated with subtype-specific monoclonal antibodies targeting the haemagglutinin proteins of H5, H7, and H10 AIVs. The assay achieved limits of detection of 0.0313, 0.0156, and 0.0625 ng/mL for recombinant HA proteins and 2[SUP]-4[/SUP], 2[SUP]-4[/SUP], and 2[SUP]-3[/SUP] haemagglutination units for viral titers of H5, H7, and H10, respectively. No cross-reactivity was observed with other AIV subtypes or respiratory pathogens, and intra- and inter-assay variation remained below 6%, demonstrating high specificity and reproducibility. Validation with 135 avian and 125 human clinical samples showed complete concordance with real-time RT-PCR results. Integration with a smartphone-based analytical platform enabled rapid readout, automated quantification, and cloud-based data sharing, providing results within 10 minutes. This intelligent UCNPs-LFA system combines ultra sensitivity, multiplexing, and field-deployable usability, representing a significant advance over conventional methods. By enabling timely and reliable detection of H5, H7, and H10 AIVs in both animal and human samples, this platform offers a practical tool for early warning, surveillance, and control of emerging zoonotic influenza, thereby contributing to global preparedness against avian influenza outbreaks.
Keywords: Avian influenza virus; Multiplex immunochromatographic assay; Smartphone; Ultrasensitive detection; Upconversion nanoparticles.
. 2025 Dec 9:2602315.
doi: 10.1080/22221751.2025.2602315. Online ahead of print. Smartphone-assisted upconversion nanoparticle assay for rapid multiplex detection of H5, H7, and H10 avian influenza viruses
Jun Zhang[SUP] 1 [/SUP], Han Wu[SUP] 1 [/SUP], Ping Wang[SUP] 1 [/SUP], Jiamin Fu[SUP] 1 [/SUP], Xinyao Zheng[SUP] 2 [/SUP], Fang Wan[SUP] 3 [/SUP], Man Hu[SUP] 2 [/SUP], Fumin Liu[SUP] 1 [/SUP], Linfang Cheng[SUP] 1 [/SUP], Hangping Yao[SUP] 1 [/SUP], Nanping Wu[SUP] 1 [/SUP], Junli Zhu[SUP] 2 [/SUP], Haibo Wu[SUP] 1 [/SUP]
Affiliations
- PMID: 41363045
- DOI: 10.1080/22221751.2025.2602315
Avian influenza viruses (AIVs) of the H5, H7, and H10 subtypes pose substantial threats to global public health owing to their high pathogenicity, cross-species transmissibility, and potential to spark epidemics. Rapid and accurate detection is essential for outbreak control and zoonotic risk mitigation. Here, we report the development of a multiplex lateral flow immunoassay (LFA) based on core-shell upconversion nanoparticles (UCNPs) conjugated with subtype-specific monoclonal antibodies targeting the haemagglutinin proteins of H5, H7, and H10 AIVs. The assay achieved limits of detection of 0.0313, 0.0156, and 0.0625 ng/mL for recombinant HA proteins and 2[SUP]-4[/SUP], 2[SUP]-4[/SUP], and 2[SUP]-3[/SUP] haemagglutination units for viral titers of H5, H7, and H10, respectively. No cross-reactivity was observed with other AIV subtypes or respiratory pathogens, and intra- and inter-assay variation remained below 6%, demonstrating high specificity and reproducibility. Validation with 135 avian and 125 human clinical samples showed complete concordance with real-time RT-PCR results. Integration with a smartphone-based analytical platform enabled rapid readout, automated quantification, and cloud-based data sharing, providing results within 10 minutes. This intelligent UCNPs-LFA system combines ultra sensitivity, multiplexing, and field-deployable usability, representing a significant advance over conventional methods. By enabling timely and reliable detection of H5, H7, and H10 AIVs in both animal and human samples, this platform offers a practical tool for early warning, surveillance, and control of emerging zoonotic influenza, thereby contributing to global preparedness against avian influenza outbreaks.
Keywords: Avian influenza virus; Multiplex immunochromatographic assay; Smartphone; Ultrasensitive detection; Upconversion nanoparticles.