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印度南部猪圆环病毒2型的分离与基因分析表明猪圆环病毒2d基因型广泛传播。

Isolation and genetic analysis of Porcine circovirus 2 in southern India evidences high circulation of Porcine circovirus 2d genotype.

作者信息

Parthiban S, Ramesh A, Karuppannan Anbu Kumar, Dhinakar Raj G, Hemalatha S, Parthiban M, Senthilkumar K, Balasubramaniyam D, Sumanth Kumar R, Ranganatha S, Ravishankar Chintu

机构信息

Department of Animal Biotechnology, Madras Veterinary College, Chennai, 600 007, India.

Vaccine Research Centre-Viral Vaccines, CAHS, MMC, Chennai, 600 051, India.

出版信息

Mol Biol Rep. 2022 Dec;49(12):11829-11846. doi: 10.1007/s11033-022-07946-5. Epub 2022 Oct 10.

Abstract

BACKGROUND

Porcine circovirus 2 is globally noted swine pathogen with multiple genotypes associated with vast clinical and subclinical outcomes. This study aimed to isolate and characterize PCV2 genotypes circulating in southern states of India.

METHODS AND RESULTS

A total of 434 field samples comprising of serum (n = 273), tissues (n = 109) and swabs (n = 52) collected from swine during 2019 to 2021 from southern states of India were screened for PCV2 by specific polymerase chain reaction (PCR) assay. Molecular prevalence of PCV2 in southern India was found to be 12.21% (n = 53). All the 53 PCV2 positive samples were further subjected to the PCR assay with designed primers targeting full length amplification of ORF2 gene of PCV2 for molecular characterization. Randomly 32 positive samples by full length PCV2-ORF2 gene PCR were sequenced for genotyping. Signature motif and phylogenetic analysis of 32 PCV2 sequences revealed 62.5% (n = 20) prevalence of PCV2d genotype followed by 21.8% (n = 7) of PCV2h or PCV2-IM1 and 15.6% (n = 5) of PCV2b genotypes. Twenty five PCR positive field samples were subjected for virus isolation in PK15 cells and characterized. Out of 25 samples processed 5 (20%) PCV2 isolates obtained in this study were confirmed by PCR and immune fluorescence assay. Molecular characterization of PK15 adapted five PCV2 isolates confirmed circulation of PCV2d, PCV2h and PCV2b genotypes in pigs under field conditions in southern India.

CONCLUSIONS

Isolation and molecular epidemiological study of PCV2 in southern states of India evidences high circulation of PCV2d genotypes in field conditions in comparison to other genotypes.

摘要

背景

猪圆环病毒2型是一种在全球范围内受到关注的猪病原体,有多种基因型,与广泛的临床和亚临床结果相关。本研究旨在分离和鉴定在印度南部各邦流行的猪圆环病毒2型基因型。

方法与结果

2019年至2021年期间,从印度南部各邦的猪身上采集了总共434份现场样本,包括血清(n = 273)、组织(n = 109)和拭子(n = 52),通过特异性聚合酶链反应(PCR)检测法对猪圆环病毒2型进行筛查。结果发现印度南部猪圆环病毒2型的分子流行率为12.21%(n = 53)。对所有53份猪圆环病毒2型阳性样本进一步进行PCR检测,使用设计的引物靶向猪圆环病毒2型ORF2基因的全长扩增以进行分子鉴定。随机选取32份通过猪圆环病毒2型全长ORF2基因PCR检测呈阳性的样本进行测序以进行基因分型。对32条猪圆环病毒2型序列的特征基序和系统发育分析显示,猪圆环病毒2d基因型的流行率为62.5%(n = 20),其次是猪圆环病毒2h或猪圆环病毒2-IM1的21.8%(n = 7)和猪圆环病毒2b基因型的15.6%(n = 5)。对25份PCR阳性现场样本在PK15细胞中进行病毒分离并鉴定。在处理的25份样本中,本研究获得的5份(20%)猪圆环病毒2型分离株通过PCR和免疫荧光检测得到确认。对适应PK15的5份猪圆环病毒2型分离株的分子鉴定证实,在印度南部的野外条件下,猪圆环病毒2d、猪圆环病毒2h和猪圆环病毒2b基因型在猪中流行。

结论

印度南部各邦猪圆环病毒2型的分离和分子流行病学研究表明,与其他基因型相比,猪圆环病毒2d基因型在野外条件下的流行率较高。

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