1. An affordable detection system based on RT-LAMP and DNA-nanoprobes for avian metapneumovirus.
- Author
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Cea-Callejo P, Arca-Lafuente S, Gomez-Lucia E, Doménech A, Biarnés M, Blanco A, Benítez L, and Madrid R
- Subjects
- Animals, Turkeys, Metal Nanoparticles chemistry, Limit of Detection, Colorimetry methods, DNA, Viral genetics, Metapneumovirus genetics, Metapneumovirus isolation & purification, Nucleic Acid Amplification Techniques methods, Nucleic Acid Amplification Techniques economics, Chickens virology, Molecular Diagnostic Techniques methods, Molecular Diagnostic Techniques economics, Paramyxoviridae Infections diagnosis, Paramyxoviridae Infections veterinary, Paramyxoviridae Infections virology, Poultry Diseases virology, Poultry Diseases diagnosis, Gold chemistry, Sensitivity and Specificity
- Abstract
Airborne animal viral pathogens can rapidly spread and become a global threat, resulting in substantial socioeconomic and health consequences. To prevent and control potential epidemic outbreaks, accurate, fast, and affordable point-of-care (POC) tests are essential. As a proof-of-concept, we have developed a molecular system based on the loop-mediated isothermal amplification (LAMP) technique for avian metapneumovirus (aMPV) detection, an airborne communicable agent mainly infecting turkeys and chickens. For this purpose, a colorimetric system was obtained by coupling the LAMP technique with specific DNA-functionalized AuNPs (gold nanoparticles). The system was validated using 50 different samples (pharyngeal swabs and tracheal tissue) collected from aMPV-infected and non-infected chickens and turkeys. Viral detection can be achieved in about 60 min with the naked eye, with 100% specificity and 87.88% sensitivity for aMPV. In summary, this novel molecular detection system allows suitable virus testing in the field, with accuracy and limit of detection (LOD) values highly close to qRT-PCR-based diagnosis. Furthermore, this system can be easily scalable to a platform for the detection of other viruses, addressing the current gap in the availability of POC tests for viral detection in poultry farming. KEY POINTS: •aMPV diagnosis using RT-LAMP is achieved with high sensitivity and specificity. •Fifty field samples have been visualized using DNA-nanoprobe validation. •The developed system is a reliable, fast, and cost-effective option for POCT., (© 2024. The Author(s).)
- Published
- 2024
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