Development of a recombinase-aided amplification assay for rapid and sensitive detection of porcine circovirus 3

Yongdong Li; Zhaorong Yu; Suli Jiao; Yuhui Liu; Hongxia Ni; Yong Wang

doi:10.1016/j.jviromet.2020.113904

Development of a recombinase-aided amplification assay for rapid and sensitive detection of porcine circovirus 3

J Virol Methods. 2020 May 26:282:113904. doi: 10.1016/j.jviromet.2020.113904. Online ahead of print.

Authors

Yongdong Li¹, Zhaorong Yu², Suli Jiao¹, Yuhui Liu¹, Hongxia Ni¹, Yong Wang³

Affiliations

¹ Municipal Key Laboratory of Virology, Ningbo Municipal Center for Disease Control and Prevention, Ningbo 315010, PR China.
² Anhui Province Key Laboratory of Veterinary Pathobiology and Disease Control, College of Animal Science and Technology, Anhui Agricultural University, Hefei 230036, PR China.
³ Anhui Province Key Laboratory of Veterinary Pathobiology and Disease Control, College of Animal Science and Technology, Anhui Agricultural University, Hefei 230036, PR China. Electronic address: wangyong119@ahau.edu.cn.

PMID: 32470487
DOI: 10.1016/j.jviromet.2020.113904

Abstract

Porcine circovirus type 3 (PCV3) is a novel member of the genus Circovirus, first detected in the United States in 2016, with subsequent reports in many countries. PCV3 infections have caused serious economic losses in the pig industry. Alternative rapid and sensitive assays for PCV3 detection are needed for clinical diagnosis, especially in laboratories not equipped with more sophisticated equipment. Here, a real-time recombinase-aided amplification assay (RAA) was developed for PCV3 detection. Specific primers and probes targeting the conserved region of the capsid gene of PCV3 were designed. The assay was performed at 39 °C for 30 min using specialized equipment. Furthermore, 36 clinical samples were used to evaluate the RAA. The analytical sensitivity of the RAA for PCV3 was 38 copies per reaction at 95% probability level, using a probit regression model. There was no cross-reactivity with other DNA viruses belonging to the Circoviridae and Parvoviridae families. The detection rate agreed with that obtained by an established real-time PCR assay with a kappa value of 1.0. Our results demonstrated that this new RAA could be used for the rapid, accurate, and sensitive detection of PCV3.

Keywords: Porcinecir covirus 3; Rapid detection; Recombinase-aided amplification assay.