Isolation and analysis of a very virulent Marek’s disease virus strain in China

Virology Journal

Hongchao Ma²  Zhiliang Wang²  Hu Shan¹  Linlin Chen¹  Jianlin Wang¹  Lijuan Zhang²  Zhenhua Gong² 

[1] Qingdao Agriculture University, Qingdao 266109, China;China Animal Healthy and Epidemic Center, Qingdao 266032, China

关键词: Immunization;    Virulence;    Oncogenic gene;    PCR;    Isolation;    Marek’s disease virus;   

Others  :  1150306
DOI  :  10.1186/1743-422X-10-155

 received in 2013-03-05, accepted in 2013-05-15,  发布年份 2013

【 摘 要 】

Background

A severe MD was broken out at a farm in Shandong, China, despite FC126 vaccination of the chickens at 1-day-old. The mortality of the flocks reached up to 38.3%. The infected chickens were found to have MD pathological changes, including enlargement of spleens, livers and kidneys, and tumors occured on organs later. Samples were collected from the chickens for diagnosis.

Methods

The collected samples were inoculated into primary duck embryo fibroblast (DEF) cells, and the MDV strain named SD2012-1 was isolated. In order to identify the isolate, amplification by PCR and sequencing of oncogenic Meq and vIL-8 gene were processed, the obtained sequences were compared with the sequences of reference strains, and SD2012-1 was used to challenge immunized SPF chickens.

Results

A very virulent MDV isolate strain, SD2012-1, was isolated from a chicken flock in Shandong Province, China, the isolate had the characteristics of very virulent MDV-1, nucleotide and deduced amino acid sequence comparisons of Meq and vIL-8 gene of SD2012-1 with those of reference strains showed SD2012-1 had high homology with MDV strains isolated from China, SD2012-1 could break through the protection provided by HVT vaccine and HVT + SB-1 vaccine immunization and caused the mortality of SPF chickens over 60%. The immune failure occured at the farm could be due to the improper selection of vaccines. SD2012-1 produced death later and the gross postmortem lesions of chickens died early and later were different.

Conclusions

MDV strain SD2012-1 isolated from Shandong Province, China was found to have the characteristics of very virulent MDV-1, which could break through the protection provided by HVT vaccine and HVT + SB-1 vaccine, the virus seemed to have a long latent period, and cause different gross postmortem lesions of chickens between chickens died early and later. A better immunization way should be chosen to prevent infection of this MDV strain in field.

【 授权许可】

   
2013 Gong et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

Figure 4.

Figure 5.

Figure 6.

【 参考文献 】

• [1]Witter RL: Increased virulence of Marek’s disease virus field isolates. Avian Dis 1997, 41:149-163.
• [2]Osterrieder N, Kamil JP, Schumacher D, Tischer BK: Trapp S. Marek’s disease virus from miasma to model. Nat Rev Microbiol 2006, 4:753-761.
• [3]Bulow VV, Biggs PM: Differentiation between strains of Marek’s disease virus and turkey herpesvirus by immunofluorescence assays. Avian Pathol 1975, 4:133-146.
• [4]Witter RL: Characteristics of Marek’s disease viruses isolated from vaccinated commercial chicken flocks: association of viral pathotype with lymphoma frequency. Avian Dis 1983, 27:113-132.
• [5]Witter RL, Calnek BW, Buscaglia C, Gimeno IM, Schat KA: Classification of Marek’s disease viruses according to pathotype: philosophy and methodology. Avian Pathol 2005, 34:75-90.
• [6]Rispens BH, van Vloten H, Mastenbroek N, Maas HJ, Schat KA: Control of Marek’s disease in the Netherlands. I. Isolation of an avirulent Marek’s disease virus (strain CVI 988) and its use in laboratory vaccination trials. Avian Dis 1972, 16:108-125.
• [7]de Boer GF, Groenendal JE, Boerrigter HM, Kok GL, Pol JM: Protective efficacy of Marek’s disease virus (MDV) CVI-988 CEF65 clone C against challenge infection with three very virulent MDV strains. Avian Dis 1986, 30:276-283.
• [8]Burgess SC, Young JR, Baaten BJ, Hunt L, Ross LN, Parcells MS, Kumar PM, Tregaskes CA, Lee LF, Davison TF: Marek’s disease is a natural model for lymphomas overexpressing Hodgkin ‘s disease antigen (CD30). Proc Natl Acad Sci U S A 2004, 101:13879-13884.
• [9]Schumacher D, Tischer BK, Teifke JP, Wink K, Osterrieder N: Generation of a permanent cell line that supports efficient growth of Marek’s disease virus (MDV) by constitutive expression of MDV glycoprotein E. J Gen Virol 2002, 83:1987-1992.
• [10]Chen M, Payne WS, Hunt H, Zhang H, Holmen SL, Dodgson JB: Inhibition of Marek’s disease virus replication by retroviral vector-based RNA interference. Virology 2008, 377:265-272.
• [11]Tian M, Zhao Y, Lin Y, Zou N, Liu C, Liu P, Cao S, Wen X, Huang Y: Comparative analysis of oncogenic genes revealed unique evolutionary features of field Marek’s disease virus prevalent in recent years in China. Virol J 2011, 8:121-131. BioMed Central Full Text
• [12]Zhang Y, Liu C, Zhang F, Shi W, Li J: Sequence analysis of the Meq gene in the predominant Marek’s disease virus strains isolated in China during 2006–2008. Virus Genes 2011, 43:353-357.
• [13]Smith EJ, Williams SM, Fadly AM: Detection of avian leukosis virus subgroup J using the polymerase chain reaction. Avian Dis 1998, 42(2):375-380.
• [14]Awad AM, Abd El-Hamid HS, Abou Rawash AA, Ibrahim HH: Detection of reticuloendotheliosis virus as a contaminant of fowl pox vaccines. Poult Sci 2010, 89:2389-2395.
• [15]Kross I, Davis PJ, Shilleto RW: Isolation of highly cytolytic MDV strains from Germany and Spain. Avian Pathol 1998, 27:313-315.
• [16]Sung HW: High virus titer in feather pulp of chickens infected with subgroup J avian leukosis virus. Avian Dis 2002, 46:517-524.
• [17]Dudnikova E, Norkina S, Vlasov A, Slobodchuk A, Lee LF, Witter RL: Evaluation of Marek’s disease field isolates by the “best fit” pathotyping assay. Avian Pathol 2007, 36:135-143.
• [18]Buscaglia C, Nervi P, Risso M: Characterization of four very virulent Argentinian strains of Marek’s disease virus and the influence of one of those isolates on synergism between Marek’s disease vaccine viruses. Avian Pathol 2004, 33:190-195.
• [19]Tan J, Cooke J, Clarke N, Tannock GA: Optimization of methods for the isolation of Marek’s disease viruses in primary chicken cell cultures. J Virol Methods 2008, 147:312-318.
• [20]Teng L, Wei P, Song Z, He J, Cui Z: Molecular epidemiological investigation of Marek’s disease virus from Guangxi, China. Arch Virol 2011, 156:203-206.
• [21]Chang K, Ohashi K, Onuma M: Diversity (polymorphism) of the Meq gene in the attenuated Marek ‘s disease virus (MDV) serotype 1 and MDV-transformed cell lines. J Vet Med Sci 2002, 64:1097-1101.
• [22]Lee LF, Wu P, Sui D, Ren D, Kamil J, Kung HJ, Witter RL: The complete unique long sequence and the overall genomic organization of the GA strain of Marek’s disease virus. Proc Natl Acad Sci U S A 2000, 97:6091-6096.
• [23]Jones D, Lee L, Liu J, Kung HJ, Tillotson JK: Marek’s disease virus encodes a basic-leucine zipper gene resembling the fos/jun oncogenes that is highly expressed inlymphoblastoid tumors. Proc Natl Acad Sci U S A 1992, 89:4042-4046.
• [24]Shamblin CE, Greene N, Arumugaswami V, Dienglewicz RL, Parcells MS: Comparative analysis of Marek’s disease virus (MDV) glycoprotein-, lytic antigen pp 38- and transformation antigen Meq-encoding genes: association of Meq mutations with MDVs of high virulence. Vet Microbiol 2004, 102:147-167.
• [25]Izumiya Y, Jang H, Ono M, Mikami T: A complete genomic DNA sequence of Marek’s disease virus type 2, strain HPRS24. Curr Top Microbiol Immunol 2001, 255:191-221.