河南省犬腺病毒Ⅱ型的流行情况调查及其分离鉴定

doi:10.11843/j.issn.0366-6964.2024.12.047

摘要/Abstract

摘要：

旨在调查犬腺病毒Ⅱ型(canine adenovirus type 2，CAV-2)在河南地区的流行特征及变异情况，于2020—2021年，从河南省8个地区随机采集790份犬粪便样品，采用PCR方法开展CAV-2病原学检测，并基于E3基因进行遗传进化分析。同时，利用MDCK细胞分离病毒，并应用负染透射电镜、细胞敏感谱试验、红细胞血凝谱试验以及理化特性分析进一步鉴定分离毒株。结果显示，CAV-2总阳性率为1.39%(11/790)，地区、年龄和免疫情况能显著影响CAV-2阳性率(P < 0.05)，其中周口市(3/40, 7.50%)感染率最高，6月龄内和免疫不全的犬易感，阳性率分别为2.69%(8/297)和1.91%(10/524)；季节和犬的性别与病毒感染无显著关系(P>0.05)。11份阳性样品测序后获得5条序列，基于CAV-2 E3基因遗传进化分析表明，4条序列与国内CAV-2 HB1株位于同一分支，1条与欧美株位于同一分支；阳性样本经蚀斑纯化后进行病毒鉴定，证实分离的病毒为CAV-2，命名为CAV2-HN21。本研究丰富了CAV-2分子流行病学数据，并成功分离出一株CAV-2毒株，为后续研究CAV-2的防治措施提供理论基础。

关键词: 河南地区, CAV-2, 流行情况调查, 病毒分离鉴定

Abstract:

In order to investigate the epidemic characteristics and variation of canine adenovirus type 2 (CAV-2) in Henan Province, a total of 790 dog fecal samples were randomly collected in eight regions of Henan Province from 2020 to 2021. The pathogen detection of CAV-2 was carried out by PCR method, and the genetic evolution analysis was carried out based on E3 gene. At the same time, MDCK cells were used to isolate the virus, and the isolated strain was further identified by negative staining transmission electron microscopy, cell sensitivity spectrum test, erythrocyte hemagglutination spectrum test and physical and chemical characteristics analysis. The results showed that the total positive rate of CAV-2 was 1.39% (11/790). Region, age and immunization status could significantly affect the positive rate of CAV-2 (P < 0.05). Among them, Zhoukou City (3/40, 7.50%) had the highest infection rate. Dogs within 6 months old and immunocompromised were susceptible, and the positive rates were 2.69% (8/297) and 1.91% (10/524), respectively. There was no significant relationship between season and sex of dogs and viral infection (P>0.05). Eleven positive samples were sequenced and 5 sequences were obtained. Based on the genetic evolution analysis of CAV-2 E3 gene, 4 sequences were located in the same branch as the domestic CAV-2 HB1 strain, and 1 sequence was located in the same branch as the European and American strains. The positive samples were identified by plaque purification and virus identification. It was confirmed that the isolated virus was CAV-2, named CAV2-HN21. This study enriched the molecular epidemiological data of CAV-2 and successfully isolated a CAV-2 strain, which provided a theoretical basis for the follow-up study of CAV-2 prevention and control measures.

Key words: Henan province, CAV-2, prevalence survey, virus isolation and identification

中图分类号:

S855.3

郭金杰, 郑力萍, 左守军, 宋鹏涛, 栗亮亮, 陈桂珍, 常志海, 代宏宇, 刘芳, 董海聚. 河南省犬腺病毒Ⅱ型的流行情况调查及其分离鉴定[J]. 畜牧兽医学报, 2024, 55(12): 5866-5872.

GUO Jinjie, ZHENG Liping, ZUO Shoujun, SONG Pengtao, LI Liangliang, CHEN Guizhen, CHANG Zhihai, DAI Hongyu, LIU Fang, DONG Haiju. Epidemiological Investigation and Isolation of Canine Adenovirus Type Ⅱ in Henan Province[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(12): 5866-5872.

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因素 Factor		阳性数/总样品数 Positive/Total	阳性率/% (95%CI) Positive rate (95%CI)	OR (95%CI)	P值 P value
地区 Region	郑州 Zhengzhou	4/150	2.67 (0.06~5.27)	1.00	0.03
	开封 Kaifeng	1/79	1.27 (-1.25~3.79)	2.137 (0.24~19.45)
	许昌 Xuchang	0/20	-	0.97 (0.95~1.00)
	焦作 Jiaozuo	1/150	0.67 (-0.65~1.98)	4.08 (0.45~36.96)
	安阳 Anyang	1/104	0.96 (-0.95~2.87)	2.82 (0.31~25.62)
	南阳 Nanyang	1/135	0.74 (-0.72~2.21)	3.67 (0.41~33.26)
	漯河 Luohe	0/112	-	0.97 (0.95~1.00)
	周口 Zhoukou	3/40	7.50 (-1.03~16.03)	0.34 (0.72~1.58)
性别 Sexuality	公犬 Male dog	5/348	1.45 (0.18~2.69)	1.00	0.93
性别 Sexuality	母犬 Female dog	6/442	1.36 (0.27~2.44)	1.06 (0.32~3.50)	0.93
年龄 Age	≤6月龄 ≤6 months old	8/297	2.69 (0.84~4.55)	1.00	0.02
年龄 Age	>6月龄 >6 months old	3/493	0.61 (-0.08~1.30)	4.521 (1.19~17.18)	0.02
免疫 Immunity	未免疫 Pre-immune	6/421	1.43 (0.29~2.56)	1.00	0.04
	未完全免疫 Not fully immune	4/103	3.88 (0.09~7.68)	0.357 (0.10~1.29)
	完全免疫 Absolute immunity	1/266	0.38 (-0.36~1.12)	3.82 (0.46~31.93)
季节 Season	春季 Spring	4/236	1.69 (0.04~3.35)	1.00	0.52
	夏季 Summer	1/115	0.87 (-0.85~2.59)	1.97 (0.22~17.79)
	秋季 Fall	1/196	0.51 (-0.50~1.52)	3.36 (0.37~30.33)
	冬季 Winter	5/243	2.06 (0.26~3.86)	0.82 (0.22~3.09)

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