| H5N1亚型禽流感病毒感染海兰白鸡模型的建立 |
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| 引用本文： | 王伟,张雅春,邓瑞坡,李越,赵颖慧,陈洪岩,孟庆文.H5N1亚型禽流感病毒感染海兰白鸡模型的建立[J].中国畜牧兽医,2015,42(3):690-695. |
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| 作者姓名： | 王伟 张雅春 邓瑞坡 李越 赵颖慧 陈洪岩 孟庆文 |
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| 作者单位： | 中国农业科学院哈尔滨兽医研究所, 兽医生物技术国家重点实验室, 哈尔滨 150001 |
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| 基金项目： | “十二五”农村领域国家科技计划课题(2011AA100305-2);中央级公益性科研院所基本科研业务费(0302014022);国家重点实验室自主研究课题(SKLVBP201429) |
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| 摘 要： | 为建立H5N1亚型禽流感病毒感染海兰白鸡模型,本研究选取1株鹅源H5N1高致病性禽流感病毒A/goose/guangdong/1/96(H5N1)(简称GD1/96),测定其对4周龄海兰白鸡的半数致死量。感染模型试验中,将30只4周龄海兰白鸡随机分成3组,每组10只,5只直接感染,5只同居,试验组设置一个重复,将病毒液稀释至104.5EID50,滴鼻、点眼各0.1mL,对照组接种PBS,感染后24h放入同居鸡;感染后连续观察14d,记录死亡时间,每天采集咽喉拭子和泄殖腔拭子;感染组和同居组第3、5天各剖解3只鸡,采集气管、肺脏、脑、脾脏、肾脏和十二指肠,进行病毒分离;qRT-PCR法分析感染组和同居组第3、5天鸡肺组织中IFN-α和TNF-α的相对表达量。结果显示,GD1/96株的鸡胚半数感染量(EID50)为10-8.167/0.1mL,对4周龄海兰白鸡的半数致死量为104.5 EID50。感染模型试验结果显示,以104.5 EID50的攻毒剂量感染海兰白鸡,感染组鸡在感染后8d全部死亡;在感染和同居3d后,各组鸡的咽喉拭子和泄殖腔拭子均可检测到病毒;感染和同居后第3、5天,各组鸡的6种组织中均可分离到高滴度的病毒;IFN-α和TNF-α在感染组和同居组的鸡肺脏组织中的表达量均显著增加(P0.05)。本试验建立了海兰白鸡的H5N1亚型禽流感病毒感染模型,为H5N1亚型禽流感病毒的致病机理及表达抗流感基因转基因鸡的研究奠定了基础。
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| 关 键 词： | 禽流感 H5N1 海兰白鸡 致病性 感染模型 |
Establishment of Model for Hy-line Variety White Chickens Infected by H5N1 Subtype Avian Influenza Virus |
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| WANG Wei;ZHANG Ya-chun;DENG Rui-po;LI Yue;ZHAO Ying-hui;CHEN Hong-yan;MENG Qing-wen.Establishment of Model for Hy-line Variety White Chickens Infected by H5N1 Subtype Avian Influenza Virus[J].China Animal Husbandry & Veterinary Medicine,2015,42(3):690-695. |
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| Authors: | WANG Wei;ZHANG Ya-chun;DENG Rui-po;LI Yue;ZHAO Ying-hui;CHEN Hong-yan;MENG Qing-wen |
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| Institution: | State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences, Harbin 150001, China |
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| Abstract: | The study was aimed to establish the suitable model for Hy-line Variety White chickens infected by H5N1 subtype avian influenza virus (AIV).The AIV used in this study was A/goose/guangdong/1/96 (H5N1) (GD1/96) virus,which was saved a goose source of highly pathogenic AIV,egg infectious dose (EID50) and median lethal dose (LD50) of 4-week-old Hy-line Variety White chickens were detected.In the infection model study,thirty 4-week-old chickens were randomly divided into three groups with 10 chickens in each group,5 direct infection,five cohabitation,the virus was diluted to 104.5 EID50,the infection chicken were inoculated with 0.1 mL to nasal,eye each,the control chicken received PBS,24 h after infecting the cohabitation chicken were added; clinical signs were assessed daily for 14 d post-infection,the time of death were record,throat swabs and cloacal swabs were collected daily,necropsy of three chickens was performed on 3 and 5 d post infection and cohabitation,the trachea,lung,brain,spleen,kidney and duodenum were collected for virus isolation.The expression of IFN-α and TNF-α in 3 and 5 d post infection and cohabitation was qualified by qRT-PCR.The results showed that the EID50 and LD50 of 4-week-old Hy-line Variety White chickens were 10-8.167/0.1 mL and 104.5 EID50,respectively.The infection model study showed that all chickens died 8 d after infection with 104.5 EID50 of the infection dosage,3 d post infection and cohabitation,high titers of AIV were detected in chicken throat swabs and cloacal swabs; 6 kinds of tissues in 3 and 5 d post infection and cohabitation were isolated to be high titer virus; the expression of IFN-α and TNF-α in lung tissue were significantly increased in infected chickens and cohabitation group (P <0.05).The model for Hy-line Variety White chickens infected by H5N1 subtype AIV was established.This study would lend some references for study of the pathogenic mechanism of H5N1 subtype AIV and generation of anti-influenza transgenic chickens against influenza virus research. |
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| Keywords: | avian influenza virus H5N1 Hy-line Variety White chicken pathogenicity infection model |
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