山羊Aichivirus C的分离鉴定及演化分析

doi:10.11843/j.issn.0366-6964.2024.08.032

Abstract

Abstract:

Aichivirus C is a newly confirmed virus associated with caprine diarrhoea. The purpose of this study was to isolate goat Aichivirus C and analyze the genomic characteristics. Nine fecal samples and 13 tissue samples from a dead kid were collected from a goat farm with an outbreak of diarrhoea in Sichuan province, and used for testing the common pathogens associated with diarrhoea. Virus was isolated from Aichivirus C positive samples using Vero cells and identified by PCR, indirect immunofluorescence and electron microscopy. The complete genomes of the isolates were amplified and analyzed. Results were as follows: 8 fecal samples with diarrhoea and 8 tissue samples from a lamb were detected as positive for Aichivirus C by RT-PCR, and no other common diarrhoea-causing pathogens were detected in the above samples. Two Aichivirus C strains were successfully isolated from positive samples using Vero cells, and the virus titers were 10^6.5TCID₅₀·mL^-1 and 10^5.9TCID₅₀·mL^-1, respectively. The nearly full-length genomic sequences of both strains were obtained, which shared 78.8%-97.4% nucleotide identity with the known goat Aichivirus C genomes from GenBank and 97.0%-97.4% nucleotide identity with the SWUN/F11/2019 isolate from China. Furthermore, the VP0, VP3 and VP1 genes were successfully amplified from 5 clinical positive samples. Phylogenetic analysis based on the genome revealed that the two isolates were clustered with the SWUN/F11/2019 strain. Unique amino acid mutations were also revealed in the VP0 and VP1 genes. In summary, two strains of goat Aichivirus C were successfully isolated and the nearly full-length genomic sequences were obtained. Phylogentic analysis demonstrated that these two strains are closely related to the SWUN/F11/2019 strain from China, with a unique molecular characteristics in the VP0 and VP1 genes. The results enrich the molecular genetic information of Aichivirus C from goats in China and laid the foundation for further studies on the pathogenicity and molecular biological characteristics of goat Aichivirus C.

Key words: goat, diarrhea, Aichivirus C, isolation, molecular characterisation

CLC Number:

S852.659.6

Yuting CHENG, ABI-Kehamo, Chen YANG, Dingzhong ZHANG, Yunxin REN, Hua YUE, Cheng TANG. Isolation and Molecular Characterization of Aichivirus C from Goats[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(8): 3612-3622.

Figures/Tables 8

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引物名称 Primer Name	上游引物(F) Forward	上游引物(R) Reverse	片段大小/bp Amplification size
1	5′-TCTTTTCCGGTGGCCA-3′	5′-TCGGCATCAGTGGTGAAAGA-3′	1 065
2	5′-CCCACCATTCCTCCCACCTT-3′	5′-AGGGTAGGCTCGCACAAAGG-3′	1 009
3	5′-CCCACCATTCCTCCCACCTT-3′	5′-CGGTAGAGCCCGTCGTAATC-3′	738
4	5′-AGTGGACTGCCCTTCAACAA-3′	5′-CACAGGTGGGCGATAGGTTT-3′	626
5	5′-AATGCGTTTGGTTCTGTTGTG-3′	5′-ACATCGGTCTCGCCATCGTA-3′	1 203
6	5′-CCAGGTTGTCCCGCTTTCAT-3′	5′-TCGGGTGAACGGCACGCAAT-3′	440
7	5′-TCTCCGCTGTCGTCGCTTAC-3′	5′-AGGGCTTGTGGGTCCGTCTT-3′	886
8	5′-CAGTCAATGCGGTGAAGAACA-3′	5′-AAACAGCACGCTCAAAGGAA-3′	1 092
9	5′-GTGCTGTTTCCCAGAACGCTCCC-3′	5′-CGAGTTTGGATTTGCGGTTG-3′	912
10	5′-GGGTGAATGTCAACCGCAAAT-3′	5′-CCCGTAAATGTGACGGGA-3′	803
11	5′-AAACTAACTGGAGATGAAAGGGTC-3′	5′-TCTCCTGCCTCAGCCTCCCT	784
12	5′-CCAGGGAGGCTGAGGCAGGAGAAT-3′	5′-AAGAACTTAATGCTCTCTAATTTTTTT-3′	153

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Isolation and Molecular Characterization of Aichivirus C from Goats

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