期刊论文详细信息
BMC Microbiology
Identification of a novel Getah virus by Virus-Discovery-cDNA random amplified polymorphic DNA (RAPD)
Fuqiang Zhang1  Quanshui Fan1  Weiguo Zhou1  Ziliang Feng1  Xiaofang Zhou1  Chaoxiong Zhang1  Yiyin Wang1  Qinghua Cui1  Jing Yu1  Wei Qiu1  Gangshan Li1  Yan Zhang2  Ying Zheng1  Tingsong Hu1 
[1] Centre for Disease Control and Prevention, Chengdu Military Region, Kunming, 650032, China;Department of Biochemistry and Molecular Biology, Fudan University Shanghai Medical College, Shanghai, 200030, China
关键词: cDNA RAPD;    Virus-Discovery;    Identification;    Getah virus;   
Others  :  1144666
DOI  :  10.1186/1471-2180-12-305
 received in 2012-04-16, accepted in 2012-12-18,  发布年份 2012
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【 摘 要 】

Background

The identification of new virus strains is important for the study of infectious disease, but current (or existing) molecular biology methods are limited since the target sequence must be known to design genome-specific PCR primers. Thus, we developed a new method for the discovery of unknown viruses based on the cDNA - random amplified polymorphic DNA (cDNA-RAPD) technique. Getah virus, belonging to the family Togaviridae in the genus Alphavirus, is a mosquito-borne enveloped RNA virus that was identified using the Virus-Discovery-cDNA RAPD (VIDISCR) method.

Results

A novel Getah virus was identified by VIDISCR from suckling mice exposed to mosquitoes (Aedes albopictus) collected in Yunnan Province, China. The non-structural protein gene, nsP3, the structural protein gene, the capsid protein gene, and the 3'-untranslated region (UTR) of the novel Getah virus isolate were cloned and sequenced. Nucleotide sequence identities of each gene were determined to be 97.1–99.3%, 94.9–99.4%, and 93.6–99.9%, respectively, when compared with the genomes of 10 other representative strains of Getah virus.

Conclusions

The VIDISCR method was able to identify known virus isolates and a novel isolate of Getah virus from infected mice. Phylogenetic analysis indicated that the YN08 isolate was more closely related to the Hebei HB0234 strain than the YN0540 strain, and more genetically distinct from the MM2021 Malaysia primitive strain.

【 授权许可】

   
2012 Hu et al.; licensee BioMed Central Ltd.

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【 参考文献 】
  • [1]Weaver SC: Host range, amplification and arboviral disease emergence. Arch Virol Suppl 2005, 19:33-44.
  • [2]Pfeffer M, Kinney RM, Kaaden OR: The alphavirus 3'-nontranslated region: size heterogeneity and arrangement of repeated sequence elements. Virology 1998, 240(1):100-108.
  • [3]Liu H, Gao X, Liang G: Newly recognized mosquito-associated viruses in mainland China, in the last two decades. Virol J 2011, 8(1):68. BioMed Central Full Text
  • [4]Kanamitsu M, Taniguchi K, Urasawa S, Ogata T, Wada Y, Wada Y, Saroso JS: Geographic distribution of arbovirus antibodies in indigenous human populations in the Indo-Australian archipelago. Am J Trop Med Hyg 1979, 28(2):351-363.
  • [5]Li XD, Qiu FX, Yang H, Rao YN, Calisher CH: Isolation of Getah virus from mosquitos collected on Hainan Island, China, and results of a serosurvey. Southeast Asian J Trop Med Public Health 1992, 23(4):730-734.
  • [6]Marchette NJ, Rudnick A, Garcia R: Alphaviruses in Peninsular Malaysia: II. Serological evidence of human infection. Southeast Asian J Trop Med Public Health 1980, 11(1):14-23.
  • [7]Allander T, Tammi MT, Eriksson M, Bjerkner A, Tiveljung-Lindell A, Andersson B: Cloning of a human parvovirus by molecular screening of respiratory tract samples. Proc Natl Acad Sci U S A 2005, 102(36):12891-12896.
  • [8]van der Hoek L, Pyrc K, Jebbink MF, Vermeulen-Oost W, Berkhout RJ, Wolthers KC, Wertheim-van Dillen PM, Kaandorp J, Spaargaren J, Berkhout B: Identification of a new human coronavirus. Nat Med 2004, 10(4):368-373.
  • [9]Williams JG, Kubelik AR, Livak KJ, Rafalski JA, Tingey SV: DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Res 1990, 18(22):6531-6535.
  • [10]Yoshida KT, Naito S, Takeda G: cDNA cloning of regeneration-specific genes in rice by differential screening of randomly amplified cDNAs using RAPD primers. Plant Cell Physiol 1994, 35(7):1003-1009.
  • [11]Yu K, Pauls KP: Optimization of the PCR program for RAPD analysis. Nucleic Acids Res 1992, 20(10):2606.
  • [12]Wen JS, Zhao WZ, Liu JW, Zhou H, Tao JP, Yan HJ, Liang Y, Zhou JJ, Jiang LF: Genomic analysis of a Chinese isolate of Getah-like virus and its phylogenetic relationship with other Alphaviruses. Virus Genes 2007, 35(3):597-603.
  • [13]George J, Raju R: Alphavirus RNA genome repair and evolution: molecular characterization of infectious sindbis virus isolates lacking a known conserved motif at the 3' end of the genome. J Virol 2000, 74(20):9776-9785.
  • [14]Hardy RW, Rice CM: Requirements at the 3' end of the sindbis virus genome for efficient synthesis of minus-strand RNA. J Virol 2005, 79(8):4630-4639.
  • [15]Kuhn RJ, Griffin DE, Zhang H, Niesters HG, Strauss JH: Attenuation of Sindbis virus neurovirulence by using defined mutations in nontranslated regions of the genome RNA. J Virol 1992, 66(12):7121-7127.
  • [16]Kuhn RJ, Hong Z, Strauss JH: Mutagenesis of the 3' nontranslated region of Sindbis virus RNA. J Virol 1990, 64(4):1465-1476.
  • [17]Raju R, Hajjou M, Hill KR, Botta V, Botta S: In vivo addition of poly(A) tail and AU-rich sequences to the 3' terminus of the Sindbis virus RNA genome: a novel 3'-end repair pathway. J Virol 1999, 73(3):2410-2419.
  • [18]Zhai YG, Wang HY, Sun XH, Fu SH, Wang HQ, Attoui H, Tang Q, Liang GD: Complete sequence characterization of isolates of Getah virus (genus Alphavirus, family Togaviridae) from China. J Gen Virol 2008, 89(Pt 6):1446-1456.
  • [19]Zhao W, Zhou G, He H: Cloning and primary analysis of 3 'end genome of two alphaviruses isolated from Hainan Province of China. Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi 2000, 14(3):213-217.
  • [20]Bryant JE, Crabtree MB, Nam VS, Yen NT, Duc HM, Miller BR: Isolation of arboviruses from mosquitoes collected in northern Vietnam. Am J Trop Med Hyg 2005, 73(2):470-473.
  • [21]Chang CY, Huang CC, Huang TS, Deng MC, Jong MH, Wang FI: Isolation and characterization of a Sagiyama virus from domestic pigs. J Vet Diagn Invest 2006, 18(2):156-161.
  • [22]Norder H, Lundstrom JO, Kozuch O, Magnius LO: Genetic relatedness of Sindbis virus strains from Europe, Middle East, and Africa. Virology 1996, 222(2):440-445.
  • [23]Turell MJ, O'Guinn ML, Wasieloski LP Jr, Dohm DJ, Lee WJ, Cho HW, Kim HC, Burkett DA, Mores CN, Coleman RE, et al.: Isolation of Japanese encephalitis and Getah viruses from mosquitoes (Diptera: Culicidae) collected near Camp Greaves, Gyonggi Province, Republic of Korea, 2000. J Med Entomol 2003, 40(4):580-584.
  • [24]Wang HQ, Liu WB, Yang DR, Liang Y, Wang JW, Zhang LS, Liu JW, Tao SJ, Lv XJ, Liang GD: Isolation and identification of arboviruses in Hebei Province. Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi 2006, 20(1):52-55.
  • [25]Hill MN, Varma MG, Mahadevan S, Meers PD: Arbovirus infections in Sarawak: observations on mosquitoes in the premonsoon period, September to December 1966. J Med Entomol 1969, 6(4):398-406.
  • [26]Faragher SG, Dalgarno L: Regions of conservation and divergence in the 3' untranslated sequences of genomic RNA from Ross River virus isolates. J Mol Biol 1986, 190(2):141-148.
  • [27]Wallner G, Mandl CW, Kunz C, Heinz FX: The flavivirus 3'-noncoding region: extensive size heterogeneity independent of evolutionary relationships among strains of tick-borne encephalitis virus. Virology 1995, 213(1):169-178.
  • [28]Boom R, Sol CJ, Salimans MM, Jansen CL, Wertheim-van Dillen PM, van der Noordaa J: Rapid and simple method for purification of nucleic acids. J Clin Microbiol 1990, 28(3):495-503.
  • [29]Zhai YG, Wang HQ, Fu SH, Liang GD: Molecular analysis on the capsid gene and 3' untranslation region of three Getah viruses isolated in China. Bing Du Xue Bao 2007, 23(4):270-275.
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