Detection of Fowl Adenovirus Serotype 4 by Recombinase Aided Amplification Combined with CRISPR/Cas13a

YIN Dongdong; GUO Hao; LAN Mengdie; YIN Lei; WANG Jieru; SHEN Xuehuai; DAI Yin; PAN Xiaocheng

doi:10.12101/j.issn.1004-390X(n).202211058

JOURNAL OF YUNNAN AGRICULTURAL UNIVERSITY（Natural Science） > 2023 > 38(5): 803-807. > DOI: 10.12101/j.issn.1004-390X(n).202211058

YIN Dongdong, GUO Hao, LAN Mengdie, et al. Detection of Fowl Adenovirus Serotype 4 by Recombinase Aided Amplification Combined with CRISPR/Cas13a[J]. JOURNAL OF YUNNAN AGRICULTURAL UNIVERSITY（Natural Science）, 2023, 38(5): 803-807. DOI: 10.12101/j.issn.1004-390X(n).202211058

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Detection of Fowl Adenovirus Serotype 4 by Recombinase Aided Amplification Combined with CRISPR/Cas13a

YIN Dongdong^{1, 2,},
GUO Hao³,
LAN Mengdie⁴,
YIN Lei^{1, 2},
WANG Jieru^{1, 2},
SHEN Xuehuai^{1, 2},
DAI Yin^{1, 2},
PAN Xiaocheng^{1, 2, ,}

1.
Livestock and Poultry Epidemic Diseases Research Center of Anhui Province, Institute of Animal Husbandry and Veterinary Science, Anhui Academy of Agricultural Science, Hefei 230001, China
2.
Anhui Provincial Key Laboratory of Livestock and Poultry Product Safety Engineering, Hefei 230001, China
3.
Animal Health Supervision Institute of Agriculture and Rural Bureau of Feixi County, Hefei 231200, China
4.
Animal Health Supervision Institute of Ningguo City, Ningguo 242300, China

More Information

Received Date: November 27, 2022
Revised Date: September 04, 2023
Accepted Date: September 05, 2023
Available Online: September 07, 2023
Published Date: October 06, 2023

Graphical Abstract

Abstract

Abstract

Purpose
To establish an efficient, sensitive and specific detection method for fowl adenovirus serotype 4 (FAdV-4).
Methods
Recombinase aided amplification (RAA) was used in combination with the clustered regularly interspaced short palindromic repeats associated protein 13a (CRISPR-Cas13a) to design RAA primers and CRISPR RNA (crRNA) targeting the conserved region of the FAdV-4 Hexon gene. The RAA technology was employed to amplify the sample nucleic acid, followed by CRISPR-Cas13a fluorescence detection. The established method was evaluated for sensitivity, specificity, and consistency compared to the qPCR control method.
Results
The method established in this study was performed at 37 ℃ with a minimum amplification copy number of 10¹ copies/μL, it was sensitive and had no cross-reactivity with avian-derived pathogens such as FAdV-1, FAdV-7, FAdV-8b, FAdV-9, FAdV-10, avian infectious laryngotracheitis virus, infectious bursal disease virus, infectious bronchitis virus, avian influenza subtype H9, newcastle disease virus. The detection rate of 30 clinical samples by this method was consistent with that of qPCR.
Conclusion
The RAA-Cas13a method is simple, sensitive and specific for the detection of FAdV-4, and can be used for rapid detection and epidemiological surveillance of FAdV-4.
- fowl adenovirus serotype 4,
- CRISPR/Cas13a,
- recombinase aided amplification,
- detection

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References (23)

References

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