Isolation，Identification and Whole Genome Sequence Analysis of Porcine Parvovirus BJ2 Strain

doi:10.15933/j.cnki.1004-3268.2023.02.015

Abstract

Abstract: To monitor the prevalence and variation of porcine parvovirus（PPV） in China，the tissue samples collected from Jiangsu，Henan and Beijing during 2020—2021 were detected by the PPV‑specific PCR method. The PPV‑positive samples in the PCR test were inoculated into porcine testicular cells（ST），and the isolated strain was identified by indirect immunofluorescence（IFA），real‑time fluorescence quantitative PCR（qPCR），and electron microscope.After that，the full genome of the strain was sequenced，which was then analyzed by MEGA 6.0 and MegAlign softwares.The isolated strain of PPV was named BJ2.At the whole genome level，the BJ2strain showed nucleotide consistency of 99.3%，99.3%，and 95.9% compared with the strains of Kresse，Challenge，and HeN1202（isolated in 2019），respectively.The VP2 gene of the BJ2 strain showed nucleotide consistency of 99.3% compared with the German strain 27a，and evolutionarily，HeN1202 was closely related to NADL‑2.Compared with the VP2 protein of the 27a strain，the amino acid sequence of BJ2 VP2 protein has two different amino acid sites.In conclusion，PPV BJ2 was evolutionarily closely related to German strain 27a.

Key words: Porcine parvovirus, Isolation and identification, VP2 gene, Sequence analysis, Antigenic drift

摘要： 为了解国内猪细小病毒（PPV）的流行、毒株变异情况，采用PPV特异性PCR方法检测2020—2021年从江苏、河南和北京3个省市采集的组织样品，将PPV阳性病料接种猪睾丸细胞（ST）进行病毒分离，利用间接免疫荧光方法（IFA）、实时荧光定量PCR（qPCR）和电镜观察对分离株进行鉴定，并测定分离株全基因组核苷酸序列，之后应用MEGA 6.0和MegAlign软件对分离株进行遗传变异分析。结果表明，成功分离到1株PPV，命名为BJ2株；全基因组核苷酸一致性分析结果显示，BJ2株与皮炎型强毒株Kresse株、Challenge株核苷酸一致性为99.3%，与实验室2019年分离株HeN1202株核苷酸一致性为95.9%；VP2基因核苷酸一致性和遗传进化分析结果显示，BJ2株与德国27a株处于同一进化分支，其核苷酸一致性为99.3%，HeN1202株与弱毒株NADL-2株亲缘关系较近；BJ2株与德国27a株相比，VP2蛋白氨基酸序列存在2个位点差异。综上，PPV BJ2株与德国27a株亲缘关系较近。

关键词: 猪细小病毒, 分离鉴定, VP2基因, 序列分析, 抗原漂移

CLC Number:

S855.3

ZHAO Wenying, ZHANG Yunjing, BAI Xiaofei, SUN Yujie, GUO Linghua, HUANG Baicheng, TIAN Kegong. Isolation，Identification and Whole Genome Sequence Analysis of Porcine Parvovirus BJ2 Strain[J]. Journal of Henan Agricultural Sciences, 2023, 52(2): 136-144.

赵文影, 张云静, 白小飞, 孙玉洁, 郭玲花, 黄柏成, 田克恭. 猪细小病毒BJ2 株的分离鉴定及全基因组序列分析[J]. 河南农业科学, 2023, 52(2): 136-144.

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