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

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:

• Fowl adenovirus serotype 4
• RAA
• CRISPR/Cas13a
• SHERLOCK; collateral cleavage; lateral flow detection

Disclosure statement

No potential conflict of interest was reported by the authors.

Additional information

Funding

This study was supported financially by the National Natural Science Foundation of China [grant number 31972642], the Major Science and Technology Special Project in Anhui Province [grant number 202003a06020012], the Anhui Province Key Research and Development Plan [grant number 202204c06020039], and the China Agriculture Research System of MOF and MARA [grant number CARS-40].