Visual detection of fowl adenovirus serotype 4 via a portable CRISPR/Cas13a-based lateral flow assay

Avian Pathol. 2023 Dec;52(6):438-445. doi: 10.1080/03079457.2023.2254253. Epub 2023 Oct 11.

Abstract

The widespread occurrence of fowl adenovirus serotype 4 (FAdV-4)-induced hepatitis-hydropericardium syndrome (HHS) has led to significant economic losses for the poultry industry. A sensitive, accurate, and practical FAdV-4 diagnostic approach is urgently required to limit the incidence of the disease. In the present study, a practical method for detecting FAdV-4 was developed using the CRISPR/Cas13a system and recombinase-aided amplification. The approach was based on 37°C isothermal detection with visible results being achieved. The detection limit of the target gene with this approach was only 101 copies/μl, making it very sensitive and specific. Clinical samples fared well when tested with the Cas13a detection method. For identifying FAdV-4, this novel detection approach was found to be sensitive, specific, and effective.RESEARCH HIGHLIGHTS First study using the CRISPR/Cas13a-based lateral flow detection assay for FAdV-4 detection.The results can be observed by the naked eye.The developed assay could provide an alternative tool for detection of FAdV-4 with minimal equipment.

Keywords: CRISPR/Cas13a; Fowl adenovirus serotype 4; RAA; SHERLOCK; collateral cleavage; lateral flow detection.

MeSH terms

  • Adenoviridae / genetics
  • Adenoviridae Infections* / diagnosis
  • Adenoviridae Infections* / veterinary
  • Animals
  • Aviadenovirus* / genetics
  • Chickens
  • Clustered Regularly Interspaced Short Palindromic Repeats
  • Poultry Diseases*
  • Serogroup