J Bacteriol Virol.  2015 Dec;45(4):328-338. 10.4167/jbv.2015.45.4.328.

Recharacterization of Morphological and Genetic Feature of Getah Virus Isolated from South Korea

Affiliations
  • 1Viral Disease Division, Animal and Plant Quarantine Agency, Gyeonggi-do, Korea. yangdk@korea.kr

Abstract

Three QIAG93 strains, QIAG9301, QIAG9302 and QIAG9303 that have been identified as Getah virus (GETV) are analyzed in this study. The morphological features of three virus isolates were observed by using electron microscopy, suggesting that the QIAG9301, QIAG9302 and QIAG9303 isolate can be classified as tentative member of Alphavirus species in the Semliki Forest complex. The full length of the structural polyprotein gene of each QIAG93 isolate (QIAG9301, QIAG9302 and QIAG9303) was determined that are identical in size, comprising 3759 nucleotides that encoded 1253 amino acids. The sequence analysis of the structural polyprotein gene, including the C, E3, E1, 6K and E2 domain, showed that each QIAG93 isolate shares >98.9% sequence identity. The phylogenetic analysis and evolutionary distance (ED) estimation based on the structural polyprotein gene sequence showed that the QIAG9301 isolate is closely related to GETV South Korea strain (99.9% sequence identity and ED value 0.001) and Chinese GETV YN0540 strain (99.3% sequence identity ED value 0.007) than other Alphavirus species analyzed in this study. Both QIAG9032 and QIAG9303 isolate exhibited genetically close relationship with Mongolian GETV LEIV17741MPR strain (at least 99.3% sequence identity and mean ED value 0.0065). Therefore, our findings will be valuable for molecular epidemiological analyses of GETV in Korea and contribute to a further study on pathogenicity of three QIAG93 isolates in animals.

Keyword

Getah virus; Genetic epidemiology

MeSH Terms

Alphavirus*
Amino Acids
Animals
Asian Continental Ancestry Group
Humans
Korea*
Microscopy, Electron
Molecular Epidemiology
Nucleotides
Sequence Analysis
Trees
Virulence
Amino Acids
Nucleotides

Figure

  • Figure 1. Electron microscopy of QIAG93 isolates. QIAG9301 was used as a representative of the three isolates. Virus-like particles in specimens prepared by direct (A) or sucrose gradient centrifugation (B). General morphological characteristics of viruses and cytopathic changes in Vero cells after virus infection for 72 h (C, D and E). Mock-infected cells were used for comparison (F). Inset C, black solid arrows indicate virus-like particles.

  • Figure 2. Amplification of the structural polyprotein gene of the GETV QIAG93 isolate with a specific primer set (see Table 1). Lanes 1 to 5, PCR products amplified using the respective GE1, GE2, GE3, GE4, and GE5 primer sets. Lane M, 100 bp DNA ladder. Use of the GE1 to GE5 primer sets resulted in amplification of products with sizes of 990, 1,030, 990, 880 and 490 bp, respectively.

  • Figure 3. Phylogenetic relationships between QIAG93 isolates and other Alphaviruses within the Semliki Forest antigenic complex. Phylogenies were assessed on the structural polyprotein gene using neighbor-joining methods. A bootstrap method with 1,000 replicates was applied in this analysis to evaluate the reliability of the inferred phylogenetic tree. In the neighbor-joining tree, the monophyletic sister groups that had bootstrap values > 70% were chosen. The GETV strains were classified into three sister groups rooted at GETV MM2021.


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Seung Heon Lee, Dong-Kun Yang, Ha-Hyun Kim, Hyun-Ye Jo, Sung-Suk Choi, In-Soo Cho
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Reference

1). Berge TO. International catalogue of arboviruses, including certain other viruses of vertebrates. Washington D. C.: US Department of Health, Education and Welfare, Public Health Service;1975. p. 278.
2). Go YY, Balasuriya UB, Lee CK. Zoonotic encephalitides caused by arboviruses: transmission and epidemiology of alphaviruses and flaviviruses. Clin Exp Vaccine Res. 2014; 3:58–77.
Article
3). Elisvberg BL, Buescher EL. Getah (MM 2021). Arbovirus catalogue card No.47, Arbovirus Information Exchange. 1963.
4). Doherty RL, Gorman BM, Whitehead RH, Carley JG. Studies of arthropod-borne virus infections in Queensland. V. Survey of antibodies to Group A arboviruses in man and other animals. Aust J Expt Biol Sci. 1966; 44:365–77.
5). Brown CM, Timoney PJ. Getah virus infection of Indian horses. Trop Anim Health Prod. 1988; 30:241–52.
6). Kamada M, Ando Y, Fukunaga Y, Kumanomido T, Imagawa H, Wada R, et al. Equine Getah virus infection: Isolation of the virus from racehorses during a enzootic in Japan. Am J Trop Med Hyg. 1980; 29:984–8.
7). Sentsui H, Kono Y. An epidemic of Getah virus infection among racehorses: isolation of the virus. Res Vet Sci. 1980; 29:157–61.
Article
8). Timoney PJ. Getah virus infection. Infectious Diseases of Livestock. Coetzer JAW and Tustin RC, editor. 2nd ed.Cape Town: Oxford University Press;2004. p. 1023–26.
9). Kumanomido T, Wada R, Kanemaru T, Kamada M, Hirasawa K, Akiyama Y. Clinical and virological observations on swine experimentally infected with Getah virus. Vet Microbiol. 1988; 16:295–301.
Article
10). Matsuyama T, Nakamura T, Isahai K, Oya A, Kobayashi M. Haruna virus, a group A arbovirus isolated from swine in Japan. Gunma J Med Sci. 1967; 16:131–4.
Article
11). Marchette NJ, Rudnick A, Garcia R. Alphaviruses in Peninsular Malaysia: II. Serological evidence of human infection. Southeast Asian J Trop Med Public Health. 1980; 11:14–23.
12). Calisher CH, Karabatsos N. Arbovirus serogroups: definition and geographic distribution. The Arboviruses: Epidemiology and Ecology. Monath TP, editor. I. Boca Raton, FL: CRC Press;1988. p. 19–57.
Article
13). Weaver SC, Dalgarno L, Frey TK, Huang HV, Kinney RM, Rice CM, et al. Family Togaviridae. Virus Taxonomy: Seventh Report of the International Committee on Taxonomy of Viruses. van Regenmortel MV, Fauguet CM, Bishop DHI, Carstens EB, Estes MK, Lemon SM, editors. San Diego: Academic Press;2000. p. 879–89.
14). Calisher CH, Shope RE, Brandt W, Casals J, Karabatsos N, Murphy FA, et al. Proposed antigenic classification of registered arboviruses I. Togaviridae, Alphavirus. Intervirology. 1980; 14:229–32.
15). Powers AM, Brault AC, Shirako Y, Strauss EG, Kang W, Strauss JH, et al. Evolutionary relationships and systematics of the alphaviruses. J Virol. 2001; 75:10118–31.
Article
16). 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; 73:470–3.
Article
17). 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; 18:156–61.
Article
18). Kanamitsu M, Taniguchi K, Urasawa S, Ogata T, Wada Y, Wada Y, et al. Geographic distribution of arbovirus antibodies in indigenous human populations in the Indo-Australian archipelago. Am J Trop Med Hyg. 1979; 28:351–63.
19). Norder H, Lundström JO, Kozuch O, Magnius LO. Genetic relatedness of Sindbis virus strains from Europe, Middle East, and Africa. Virology. 1996; 222:440–5.
Article
20). Turell MJ, O'Guinn ML, Wasieloski LP Jr, Dohm DJ, Lee WJ, Cho HW, 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; 40:580–4.
21). Tajima S, Kotaki A, Yagasaki K, Taniwaki T, Moi ML, Nakayama E, et al. Identification and amplification of Japanese encephalitis virus and Getah virus propagated from a single porcine serum sample: a case of coinfection. Arch Virol. 2014; 159:2969–75.
Article
22). Pfeffer M, Kinney RM, Kaaden OR. The Alphavirus 3′-nontranslated region: size heterogeneity and arrangement of repeated sequence elements. Virology. 1998; 240:100–8.
23). Griffin DE. Alphaviruses. Knipe DM, Howley PM, editors. Fields' Virology. 4th ed.New York: Lippincott, Williams and Wilkins;2001. p. 917–62.
24). Calisher CH, Walton TE. Getah virus infections. Virus Infections of Equines. Studdert MK, editor. Amsterdam: Elsevier;1996. p. 157–65.
25). Fukunaga Y, Kumanomido T, Kamada M. Getah virus as an equine pathogen. Vet Clin North Am Equine Pract. 2000; 16:605–17.
Article
26). Jo HY, Yang DK, Kim HH, Choi SS, Kang KS, Yang SJ, et al. Sero-surveillance of Getah virus among Thoroughbred horses in Korea. J Bacteriol Virol. 2015; 45:235–41.
Article
27). Colombet J, Sime-Ngando T. Use of PEG, Polyethylene glycol, to characterize the diversity of environmental viruses. Microb Ecol. 2012; 58:728–36.
28). Vajda BP. Concentration and purification of viruses and bacteriophages with polyethylene glycol. Folia Microbiol (Praha). 1978; 23:88–96.
Article
29). Jeanmougin F, Thompson JD, Gouy M, Higgins DG, Gibson TJ. Multiple sequence alignment with Clustal X. Trends Biochem Sci. 1998; 23:403–5.
Article
30). Thompson JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins DG. The CLUSTAL_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res. 1997; 25:4876–82.
Article
31). Hyypiä T, Hovi T, Knowles NJ, Stanway G. Classifiproperties. J Gen Virol. 1997; 78:1–11.
32). Acheson NH, Tamm I. Replication of Semliki Forest virus: an electron microscopic study. Virology. 1967; 32:128–43.
Article
33). Grimley PM, Berezesky IK, Friedman RM. Cytoplasmic structures associated with an arbovirus infection: loci of viral ribonucleic acid synthesis. J Virol. 1968; 2:1326–38.
Article
34). Fazakerley JK. Pathogenesis of Semliki Forest virus encephalitis. J Neurovirol. 2002; 8:66–74.
Article
35). Dietzgen RG, Kuzmin IV. Taxonomy of rhabdoviruses. Rhabdoviruses: Molecular Taxonomy, Evolution, Genomics, Ecology, Host-Vector Interactions, Cytopathology and Control. Dietzgen RG, Kuzmin IV, editors. Norfolk: Caister Academic Press;2012. p. 13–22.
36). Dietzschold B, Wunner WH, Wiktor TJ, Lopes AD, Lafon M, Smith CL, et al. Characterization of an antigenic determinant of the glycoprotein that correlates with pathogenicity of rabies virus. Proc Natl Acad Sci U S A. 1983; 80:70–4.
Article
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