Abstract
Waterfowl parvovirus, duck enteritis virus and goose astrovirus have become serious pathogens in waterfowl farming. Co-infections occasionally occur, and as a result, it is much harder to rapidly and simultaneously identify several pathogens using conventional PCR. According to the characteristics of the goose parvovirus (GPV) and muscovy duck parvovirus (MDPV) genome sequences, a universal PCR primer was designed using Rep1 as the target gene. The specific detection primers were designed based on the specific conserved regions of UL54 of the duck enteritis virus (DEV) gene and ORF1a of the goose astrovirus (GAstV) gene. The PCR reaction system and conditions were optimized, and the optimal annealing temperature was found to be 56.2 ℃. The volume ratio of the GPV–MDPV, GAstV and DEV primers (20 μM) was 1:4:5. The established multiplex PCR detection method can simultaneously detect GPV, MDPV, DEV and GAstV within one reaction, and be negative for duck Tembusu virus, muscovy duck reovirus, duck hepatitis A virus type 3 and duck circovirus. The method with excellent sensitivity, specificity and repeatability was successfully applied to clinical samples, it is a useful platform for identifing co-infections of GPV, MDPV, DEV and GAstV in waterfowl.
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We thank International Science Editing (http://www.internationalscienceediting.com) for editing this manuscript.
Funding
This study was supported financially by the Major Science and Technology Special Project in Anhui Province (No. 202003a06020012), the major special project of science and technology in Anhui Province (No. 201903a06020005), the Anhui Academy of Agricultural Sciences Platform Project (No. 2021YL065), and the Anhui Province Poultry Industry Technology System (No. AHCYJSTX-06).
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Dai, Y., Li, M., Hu, X. et al. Development and application of a multiplex PCR method for simultaneous detection of waterfowl parvovirus, duck enteritis virus and goose astrovirus. 3 Biotech 12, 205 (2022). https://doi.org/10.1007/s13205-022-03238-8
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DOI: https://doi.org/10.1007/s13205-022-03238-8