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

ABSTRACTThe 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 4RAACRISPR/Cas13aSHERLOCK; collateral cleavage; lateral flow detection Disclosure statementNo potential conflict of interest was reported by the authors.Additional informationFundingThis 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].