A paper-based loop-mediated isothermal amplification assay for highly pathogenic avian influenza
Abstract Avian influenza outbreaks have had significant economic and public health consequences worldwide. Therefore, prompt, reliable, and cost-effective diagnostic devices are crucial for scrutinizing and confining highly pathogenic avian influenza viruses (HPAIVs). Our study introduced and evalua...
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Nature Portfolio
2025-04-01
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| Series: | Scientific Reports |
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| Online Access: | https://doi.org/10.1038/s41598-025-95452-6 |
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| author | Mohamed Kamel Josiah Levi Davidson Jenna M. Schober Gregory S. Fraley Mohit S. Verma |
| author_facet | Mohamed Kamel Josiah Levi Davidson Jenna M. Schober Gregory S. Fraley Mohit S. Verma |
| author_sort | Mohamed Kamel |
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| description | Abstract Avian influenza outbreaks have had significant economic and public health consequences worldwide. Therefore, prompt, reliable, and cost-effective diagnostic devices are crucial for scrutinizing and confining highly pathogenic avian influenza viruses (HPAIVs). Our study introduced and evaluated a novel paper-based loop-mediated isothermal amplification (LAMP) test for diagnosing the H5 subtype of the avian influenza virus (AIV). We meticulously designed and screened LAMP primers targeting the H5-haemagglutinin (H5-HA) gene of AIV and fine-tuned the paper-based detection assay for best performance. The paper-based LAMP assay demonstrated a detection limit of 500 copies per reaction (25 copies/µl). Additionally, the assay exhibited no cross-reactivity with common bovine and avian pathogens, confirming its specificity. Spiking experiments revealed that the assay could accurately detect 1000 copies of synthetic HPAIV RNA (per reaction) when spiked into oropharyngeal swab samples, achieving 100% analytical sensitivity, specificity, and accuracy. This inexpensive, user-friendly point-of-need diagnostic tool holds great promise, especially in resource-limited settings. It only requires a water bath for incubation and enables visual detection of results without special equipment. Overall, the paper-based LAMP assay provides a promising method for rapidly and reliably detecting the H5 subtype of AIV, contributing to improved surveillance and early intervention strategies. |
| format | Article |
| id | doaj-art-901ec32e4f0b440faac70982b4b17e69 |
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| issn | 2045-2322 |
| language | English |
| publishDate | 2025-04-01 |
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| spelling | doaj-art-901ec32e4f0b440faac70982b4b17e692025-08-20T02:11:41ZengNature PortfolioScientific Reports2045-23222025-04-0115111310.1038/s41598-025-95452-6A paper-based loop-mediated isothermal amplification assay for highly pathogenic avian influenzaMohamed Kamel0Josiah Levi Davidson1Jenna M. Schober2Gregory S. Fraley3Mohit S. Verma4Department of Agricultural and Biological Engineering, Purdue UniversityDepartment of Agricultural and Biological Engineering, Purdue UniversityDepartment of Animal Sciences, Purdue UniversityDepartment of Animal Sciences, Purdue UniversityDepartment of Agricultural and Biological Engineering, Purdue UniversityAbstract Avian influenza outbreaks have had significant economic and public health consequences worldwide. Therefore, prompt, reliable, and cost-effective diagnostic devices are crucial for scrutinizing and confining highly pathogenic avian influenza viruses (HPAIVs). Our study introduced and evaluated a novel paper-based loop-mediated isothermal amplification (LAMP) test for diagnosing the H5 subtype of the avian influenza virus (AIV). We meticulously designed and screened LAMP primers targeting the H5-haemagglutinin (H5-HA) gene of AIV and fine-tuned the paper-based detection assay for best performance. The paper-based LAMP assay demonstrated a detection limit of 500 copies per reaction (25 copies/µl). Additionally, the assay exhibited no cross-reactivity with common bovine and avian pathogens, confirming its specificity. Spiking experiments revealed that the assay could accurately detect 1000 copies of synthetic HPAIV RNA (per reaction) when spiked into oropharyngeal swab samples, achieving 100% analytical sensitivity, specificity, and accuracy. This inexpensive, user-friendly point-of-need diagnostic tool holds great promise, especially in resource-limited settings. It only requires a water bath for incubation and enables visual detection of results without special equipment. Overall, the paper-based LAMP assay provides a promising method for rapidly and reliably detecting the H5 subtype of AIV, contributing to improved surveillance and early intervention strategies.https://doi.org/10.1038/s41598-025-95452-6Influenza A virusesLoop-mediated isothermal amplificationDiagnosticsPoint-of-careMicrofluidic paper-based analytical devicesµPADs |
| spellingShingle | Mohamed Kamel Josiah Levi Davidson Jenna M. Schober Gregory S. Fraley Mohit S. Verma A paper-based loop-mediated isothermal amplification assay for highly pathogenic avian influenza Scientific Reports Influenza A viruses Loop-mediated isothermal amplification Diagnostics Point-of-care Microfluidic paper-based analytical devices µPADs |
| title | A paper-based loop-mediated isothermal amplification assay for highly pathogenic avian influenza |
| title_full | A paper-based loop-mediated isothermal amplification assay for highly pathogenic avian influenza |
| title_fullStr | A paper-based loop-mediated isothermal amplification assay for highly pathogenic avian influenza |
| title_full_unstemmed | A paper-based loop-mediated isothermal amplification assay for highly pathogenic avian influenza |
| title_short | A paper-based loop-mediated isothermal amplification assay for highly pathogenic avian influenza |
| title_sort | paper based loop mediated isothermal amplification assay for highly pathogenic avian influenza |
| topic | Influenza A viruses Loop-mediated isothermal amplification Diagnostics Point-of-care Microfluidic paper-based analytical devices µPADs |
| url | https://doi.org/10.1038/s41598-025-95452-6 |
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