Isolation，identification and phylogenetic analysis of a ranavirus isolated from<italic style="font-style: italic"> Rana grylio</italic>

TIAN Silu; YUAN Yu; XU Le; OUYANG Ping; CHEN Defang; HUANG Xiaoli; GENG Yi

doi:10.13300/j.cnki.hnlkxb.2025.02.026

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Isolation，identification and phylogenetic analysis of a ranavirus isolated from Rana grylio

Animal science·Veterinary medicine | 更新时间：2026-11-05

- Isolation，identification and phylogenetic analysis of a ranavirus isolated from Rana grylio
- Journal of Huazhong Agricultural University Vol. 44, Issue 2, Pages: 258-264(2025)
- 作者机构：
  
  1.四川农业大学动物医学院，成都 611130
  2.四川农业大学动物科技学院，成都 611130
- 作者简介：
- 基金信息：
- DOI：10.13300/j.cnki.hnlkxb.2025.02.026
  CLC： Q943
- Received：05 February 2024，
  
  Published：06 March 2025
- 稿件说明：
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田思璐，袁羽，徐乐，等.1株沼泽绿蛙源蛙病毒的分离鉴定及系统发育分析［J］.华中农业大学学报，2025，44（2）：258‐264.

TIAN Silu,YUAN Yu,XU Le,et al.Isolation，identification and phylogenetic analysis of a ranavirus isolated from Rana grylio[J].Journal of Huazhong Agricultural University,2025,44(02):258-264.
田思璐，袁羽，徐乐，等.1株沼泽绿蛙源蛙病毒的分离鉴定及系统发育分析［J］.华中农业大学学报，2025，44（2）：258‐264. DOI： 10.13300/j.cnki.hnlkxb.2025.02.026.

TIAN Silu,YUAN Yu,XU Le,et al.Isolation，identification and phylogenetic analysis of a ranavirus isolated from Rana grylio[J].Journal of Huazhong Agricultural University,2025,44(02):258-264. DOI： 10.13300/j.cnki.hnlkxb.2025.02.026.

摘要

为探究四川某养殖场沼泽绿蛙（

Rana grylio

）暴发性高死亡率疾病的病因，对病蛙进行病理学检查和病原分离，并结合电镜观察、PCR检测和系统发育分析对分离病原进行鉴定。患病蛙主要病症为皮肤溃疡与四肢肿胀

；剖检见肝、脾与肾肿大。组织病理学观察见肝、脾、肾与肠道等组织器官发生变性、坏死与炎症反应。接种病料的鲤鱼上皮瘤细胞25 ℃培养 4 d呈典型细胞病变效应，TCID

为10

4.12

/0.1 mL。电镜观察到大量正六边形、对角线直径约165 nm的病毒粒子呈晶格状排列。对内脏组织及接种细胞进行蛙病毒主要衣壳蛋白（major capsid protein，MCP）基因的特异性PCR检测，结果均为阳性。基于

MCP

基因全序列的同源性和遗传进化分析显示，分离病毒与蛙病毒属病毒相似性在99%以上，且属于FV3类群。以上结果表明分离病毒（TSL210813）为蛙病毒，是本次沼泽绿蛙疫病的病因。

Abstract

To explore the etiology of a serious infectious disease affecting

Rana grylio

at a farm in Sichuan Province，pathological examinations and pathogen isolation were conducted on the diseased frogs.The isolated pathogen was identified using transmission electron microscopy，PCR detection，and phylogenetic analysis.External clinical signs included limb swelling and ulcerations on the body surface.The dead frog exhibited an enlarged liver，spleen and kidney.Histopathological observations revealed that the liver，spleen，kidney and other organs exhibited damage characterized by significant degeneration and necrosis foci.The carp epithelioma papulosum cyprini （EPC） cell line，inoculated with diseased samples at 25 ℃，demonstrated typical cytopathic effects （CPE） after four days，including rounding and shedding，with a TCID

of 10

4.12

/0.1 mL.Transmission electron microscopic observation showed that the virus exhibited a regular hexagonal shape with a crystalline array of its capsid，and the diagonal diameter measured approximately 165 nm.PCR by using specific primers for the major capsid protein （

MCP

） gene was performed on the internal tissues of infected frogs and on EPC cells with CPE，yielding positive results.Phylogenetic analysis based on

MCP

gene sequences indicated that the sequences of isolated virus shared over 99% identity with ranavirus and was classed as FV3-like.This study confirmed that the isolated virus，designated TSL210813，is a ranavirus，responsible for the disease observed in

R. grylio

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references

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Isolation，identification and phylogenetic analysis of a ranavirus isolated from Rana grylio

DOI：10.13300/j.cnki.hnlkxb.2025.02.026

摘要

Abstract

关键词

Keywords

references