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Isolation and pathogenicity of porcine circovirus type 2 in mice from Guangxi province, China

BACKGROUND: Porcine circovirus type 2 (PCV2), a member of the genus Circovirus and family Circoviridae, is a closed, small, circular, and single-stranded DNA virus, and it is a crucial swine pathogen of porcine circovirus-associated diseases (PCVADs). PCV2 was first detected in PK-15(ATCC-CCL) cells...

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Autores principales: Jiao, Qiulin, Yang, Liuyue, Liu, Xiangzu, Wen, Yanwen, Tian, Linxing, Qian, Ping, Chen, Huanchun, Li, Xiangmin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10466715/
https://www.ncbi.nlm.nih.gov/pubmed/37644571
http://dx.doi.org/10.1186/s12985-023-02161-5
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author Jiao, Qiulin
Yang, Liuyue
Liu, Xiangzu
Wen, Yanwen
Tian, Linxing
Qian, Ping
Chen, Huanchun
Li, Xiangmin
author_facet Jiao, Qiulin
Yang, Liuyue
Liu, Xiangzu
Wen, Yanwen
Tian, Linxing
Qian, Ping
Chen, Huanchun
Li, Xiangmin
author_sort Jiao, Qiulin
collection PubMed
description BACKGROUND: Porcine circovirus type 2 (PCV2), a member of the genus Circovirus and family Circoviridae, is a closed, small, circular, and single-stranded DNA virus, and it is a crucial swine pathogen of porcine circovirus-associated diseases (PCVADs). PCV2 was first detected in PK-15(ATCC-CCL) cells in 1974, which has caused significant economic loss to the swine industry throughout the world. And the first case of PCV2 was reported in China in 2000. At present, PCV2d is the main genotype circulating widely in China. METHODS: Lymph samples were obtained from piglets with emaciation and respiratory disease in Guangxi province, China. The main pathogens were detected via PCR from lymph samples, and then PCV2-single positive samples were used to inoculate with PK-15 cells. After successive generations, the isolate was subsequently identified by polymerase chain reaction (PCR), immunofluorescence assay (IFA), Western blot (WB), and transmission electron microscopic (TEM). The full-length genome and genetic characterization of isolates were analyzed by Sanger sequencing. The TCID(50) of the PCV2-GX-6 was determined by IFA, and the pathogenicity of PCV2 in BALB/c mice was analyzed via the mouse model. RESULTS: The isolates were successfully isolated from clinical samples. The complete genome of PCV2-GX-4, PCV2-GX-6, PCV2-GX-7, PCV2-GX-11 and PCV2-GX-16 have been amplified, sequenced, and deposited in GenBank (accession no.: OR133747, OQ803314, OR133748, OR133749, OR133750). Homology and phylogenetic analysis with reference strains showed that the isolates belonged to the PCV2d genotype. The PCV2-GX-6 could be stably passaged more than 30 times in PK-15 cells. PCV2-GX-6 was identified by PCR, IFA, WB and TEM. The results of homology showed that PCV2-GX-6 was closely related to the reference strains PCV2-JS17-8 (GenBank accession no.: MH211363). Pathogenicity studies in mice have shown that PCV2-GX-6 can lead to growth inhibition of mice. Meanwhile PCV2-GX-6 caused the typical lesions of spleen, lung and kidney. The results of qPCR showed that PCV2 can effectively proliferate in the liver, spleen, lung, and kidney. CONCLUSION: PCV2-GX-6 can successfully infect BLAB/c mice, effectively proliferate in major organs, and possessed high pathogenicity. In conclusion, combined with the genotype and pathogenicity of PCV2d currently prevalent, PCV2-GX-6 can be used as a candidate vaccine strain. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12985-023-02161-5.
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spelling pubmed-104667152023-08-31 Isolation and pathogenicity of porcine circovirus type 2 in mice from Guangxi province, China Jiao, Qiulin Yang, Liuyue Liu, Xiangzu Wen, Yanwen Tian, Linxing Qian, Ping Chen, Huanchun Li, Xiangmin Virol J Research BACKGROUND: Porcine circovirus type 2 (PCV2), a member of the genus Circovirus and family Circoviridae, is a closed, small, circular, and single-stranded DNA virus, and it is a crucial swine pathogen of porcine circovirus-associated diseases (PCVADs). PCV2 was first detected in PK-15(ATCC-CCL) cells in 1974, which has caused significant economic loss to the swine industry throughout the world. And the first case of PCV2 was reported in China in 2000. At present, PCV2d is the main genotype circulating widely in China. METHODS: Lymph samples were obtained from piglets with emaciation and respiratory disease in Guangxi province, China. The main pathogens were detected via PCR from lymph samples, and then PCV2-single positive samples were used to inoculate with PK-15 cells. After successive generations, the isolate was subsequently identified by polymerase chain reaction (PCR), immunofluorescence assay (IFA), Western blot (WB), and transmission electron microscopic (TEM). The full-length genome and genetic characterization of isolates were analyzed by Sanger sequencing. The TCID(50) of the PCV2-GX-6 was determined by IFA, and the pathogenicity of PCV2 in BALB/c mice was analyzed via the mouse model. RESULTS: The isolates were successfully isolated from clinical samples. The complete genome of PCV2-GX-4, PCV2-GX-6, PCV2-GX-7, PCV2-GX-11 and PCV2-GX-16 have been amplified, sequenced, and deposited in GenBank (accession no.: OR133747, OQ803314, OR133748, OR133749, OR133750). Homology and phylogenetic analysis with reference strains showed that the isolates belonged to the PCV2d genotype. The PCV2-GX-6 could be stably passaged more than 30 times in PK-15 cells. PCV2-GX-6 was identified by PCR, IFA, WB and TEM. The results of homology showed that PCV2-GX-6 was closely related to the reference strains PCV2-JS17-8 (GenBank accession no.: MH211363). Pathogenicity studies in mice have shown that PCV2-GX-6 can lead to growth inhibition of mice. Meanwhile PCV2-GX-6 caused the typical lesions of spleen, lung and kidney. The results of qPCR showed that PCV2 can effectively proliferate in the liver, spleen, lung, and kidney. CONCLUSION: PCV2-GX-6 can successfully infect BLAB/c mice, effectively proliferate in major organs, and possessed high pathogenicity. In conclusion, combined with the genotype and pathogenicity of PCV2d currently prevalent, PCV2-GX-6 can be used as a candidate vaccine strain. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12985-023-02161-5. BioMed Central 2023-08-29 /pmc/articles/PMC10466715/ /pubmed/37644571 http://dx.doi.org/10.1186/s12985-023-02161-5 Text en © The Author(s) 2023 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Research
Jiao, Qiulin
Yang, Liuyue
Liu, Xiangzu
Wen, Yanwen
Tian, Linxing
Qian, Ping
Chen, Huanchun
Li, Xiangmin
Isolation and pathogenicity of porcine circovirus type 2 in mice from Guangxi province, China
title Isolation and pathogenicity of porcine circovirus type 2 in mice from Guangxi province, China
title_full Isolation and pathogenicity of porcine circovirus type 2 in mice from Guangxi province, China
title_fullStr Isolation and pathogenicity of porcine circovirus type 2 in mice from Guangxi province, China
title_full_unstemmed Isolation and pathogenicity of porcine circovirus type 2 in mice from Guangxi province, China
title_short Isolation and pathogenicity of porcine circovirus type 2 in mice from Guangxi province, China
title_sort isolation and pathogenicity of porcine circovirus type 2 in mice from guangxi province, china
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10466715/
https://www.ncbi.nlm.nih.gov/pubmed/37644571
http://dx.doi.org/10.1186/s12985-023-02161-5
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