An affordable detection system based on RT-LAMP and DNA-nanoprobes for avian metapneumovirus

Appl Microbiol Biotechnol. 2024 Jul 10;108(1):414. doi: 10.1007/s00253-024-13243-x.

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.

Keywords: Avian metapneumovirus (aMPV); Molecular detection; Nanoprobes; Point of Care (POC) test; Reverse transcription loop mediated isothermal amplification (RT-LAMP).

MeSH terms

  • Animals
  • Chickens* / virology
  • Colorimetry / methods
  • DNA, Viral / genetics
  • Gold* / chemistry
  • Limit of Detection
  • Metal Nanoparticles / chemistry
  • Metapneumovirus* / genetics
  • Metapneumovirus* / isolation & purification
  • Molecular Diagnostic Techniques* / economics
  • Molecular Diagnostic Techniques* / methods
  • Nucleic Acid Amplification Techniques* / economics
  • Nucleic Acid Amplification Techniques* / methods
  • Paramyxoviridae Infections* / diagnosis
  • Paramyxoviridae Infections* / veterinary
  • Paramyxoviridae Infections* / virology
  • Poultry Diseases* / diagnosis
  • Poultry Diseases* / virology
  • Sensitivity and Specificity*
  • Turkeys

Substances

  • Gold
  • DNA, Viral

Supplementary concepts

  • LAMP assay