1例鸡源血清4型禽腺病毒感染的实验室诊断
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摘要
本研究从广州番禺某鸡场送检病死鸡中分离获得1株疑似Ⅰ群禽腺病毒。病鸡剖检以心包积液、肝脏呈土黄色并伴有出血点,肾脏肿胀、出血为主要特征。实验室通过血凝试验、PCR鉴定、序列测定及遗传进化分析、动物回归试验、病理组织学观察等进行一系列检测。结果显示,该毒株不能凝集鸡红细胞,根据FAdV的六邻体(Hexon)基因设计引物,进行PCR扩增和单克隆测序,PCR扩增出分子量约为550 bp的特异性条带,通过序列测定和遗传进化分析表明,该分离病毒株与FAdV-4的相似性最高。同时用该分离毒株的鸡胚尿囊液感染28日龄雏鸡,试验鸡可复制出与临床相似病例,病理组织学观察可见,心肌细胞有炎性细胞浸润,肝内有嗜碱性包涵体等病变特征。综上,分离株B13为Ⅰ群FAdV-4。
In this study, we isolated a suspected group I fowl adenovirus(FAdV) from a chicken inspected at a chicken farm in Panyu, Guangzhou. The diseased chickens were dissected with pericardial effusion, yellowish liver with bleeding points, and enlarged kidneys and bleeding as the main features.The laboratory conducts a series of tests by hemagglutination test, PCR identification, sequence phylogenetic analysis, case replication, and histopathological observation.The results showed that the strain could not agglutinate chicken red blood cells.According to the Hexon gene of FAdV, we designed primers for PCR amplification and monoclonal sequencing. PCR was used to generate specific bands with a molecular weight of approximately 550 bp.Sequence analysis and phylogenetic analysis showed that the isolate had the highest similarity to FAdV-4.At the same time, the chicken embryo allantoic fluid of the isolated strain was used to infect 4-week-old chicks, and the challenged chicken reproduced similar clinical cases.Histopathological observation revealed that there were inflammatory cell infiltration in cardiomyocytes and basophilic inclusion bodies in the liver.In summary, the isolated virus is a group I FAdV-4.
In this study, we isolated a suspected group I fowl adenovirus(FAdV) from a chicken inspected at a chicken farm in Panyu, Guangzhou. The diseased chickens were dissected with pericardial effusion, yellowish liver with bleeding points, and enlarged kidneys and bleeding as the main features.The laboratory conducts a series of tests by hemagglutination test, PCR identification, sequence phylogenetic analysis, case replication, and histopathological observation.The results showed that the strain could not agglutinate chicken red blood cells.According to the Hexon gene of FAdV, we designed primers for PCR amplification and monoclonal sequencing. PCR was used to generate specific bands with a molecular weight of approximately 550 bp.Sequence analysis and phylogenetic analysis showed that the isolate had the highest similarity to FAdV-4.At the same time, the chicken embryo allantoic fluid of the isolated strain was used to infect 4-week-old chicks, and the challenged chicken reproduced similar clinical cases.Histopathological observation revealed that there were inflammatory cell infiltration in cardiomyocytes and basophilic inclusion bodies in the liver.In summary, the isolated virus is a group I FAdV-4.
引文
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[4] Jadhao S J,Deepak J N,Kataria J M,et al.Characterisation of fowl adenoviruses from chickens affected with infectious hydropericardium during 1994-1998 in India[J].Indian Journal of Experimental Biology,2003,41(4):321- 327.
[5] Li H,Wang J,Qiu L,et al.Fowl adenovirus species C serotype 4 is attributed to the emergence of hepatitis-hydropericardium syndrome in chickens in China[J].Infection Genetics & Evolution Journal of Molecular Epidemiology & Evolutionary Genetics in Infectious Diseases,2016,45:230- 241.
[6] 罗玲,赵康,张媛,等.鸡心包积水综合征病原的分离与鉴定[J].中国家禽,2016,38(6):56- 58.
[7] Ye J,Liang G,Zhang J,et al.Outbreaks of serotype 4 fowl adenovirus with novel genotype,China[J].Emerging Microbes & Infections,2016,5(5):e50.
[8] Saifuddin M,Wilks C R.Effects of fowl adenovirus infection on the immune system of chickens[J].Journal of Comparative Pathology,1992,107(3):285.
[9] Mcferran J B.Adenoviruses of Vertebrate Animals[M]// Vertebrate Animal and Related Viruses:DNA Viruses.Elsevier Inc.1981:101- 165.
[10] Mcferran J B,Adair B M.Avian adenoviruses—a review[J].Avian Pathology,1977,6(3):189- 217.
[2] Zs??K L,Kisary J.Characterisation of adenoviruses isolated from geese[J].Avian Pathology Journal of the W.v.p.a,1984,13(2):253- 264.
[3] Shah M S,Ashraf A,Khan M I,et al.Molecular cloning,expression and characterization of 100K gene of fowl adenovirus-4 for prevention and control ofhydropericardium syndrome[J].Biologicals Journal of the International Association of Biological Standardization,2016,44(1):19- 23.
[4] Jadhao S J,Deepak J N,Kataria J M,et al.Characterisation of fowl adenoviruses from chickens affected with infectious hydropericardium during 1994-1998 in India[J].Indian Journal of Experimental Biology,2003,41(4):321- 327.
[5] Li H,Wang J,Qiu L,et al.Fowl adenovirus species C serotype 4 is attributed to the emergence of hepatitis-hydropericardium syndrome in chickens in China[J].Infection Genetics & Evolution Journal of Molecular Epidemiology & Evolutionary Genetics in Infectious Diseases,2016,45:230- 241.
[6] 罗玲,赵康,张媛,等.鸡心包积水综合征病原的分离与鉴定[J].中国家禽,2016,38(6):56- 58.
[7] Ye J,Liang G,Zhang J,et al.Outbreaks of serotype 4 fowl adenovirus with novel genotype,China[J].Emerging Microbes & Infections,2016,5(5):e50.
[8] Saifuddin M,Wilks C R.Effects of fowl adenovirus infection on the immune system of chickens[J].Journal of Comparative Pathology,1992,107(3):285.
[9] Mcferran J B.Adenoviruses of Vertebrate Animals[M]// Vertebrate Animal and Related Viruses:DNA Viruses.Elsevier Inc.1981:101- 165.
[10] Mcferran J B,Adair B M.Avian adenoviruses—a review[J].Avian Pathology,1977,6(3):189- 217.