Abstract
Getah virus (GETV), a mosquito-borne virus that mainly infects horses and pigs, has emerged and spread in China. We developed a highly specific and reproducible TaqMan probe-based quantitative reverse transcription PCR (RT-qPCR) assay targeting the non-structural protein 1 of GETV, whose detection limit is 25.5 copies/µL, which is 100-fold higher than that of conventional RT-PCR. RT-qPCR was used to detect GETV RNA in mosquito and animal clinical samples, showing that the accuracy of RT-qPCR was higher than that of conventional RT-PCR. The newly developed RT-qPCR assay may be a useful alternative tool for rapid, simple and specific diagnosis of GETV infection.
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References
Scherer WF, Funkenbusch M, Buescher EL, Izumi T (1962) Sagiyama virus, a new group A arthropod-borne virus from Japan. I. Isolation, immunologic classification, and ecologic observations. Am J Trop Med Hyg 11:255–268
Bannai H, Ochi A, Nemoto M, Tsujimura K, Yamanaka T, Kondo T (2016) A 2015 outbreak of Getah virus infection occurring among Japanese racehorses sequentially to an outbreak in 2014 at the same site. BMC Vet Res 12:98. https://doi.org/10.1186/s12917-016-0741-5
Li XD, Qiu FX, Yang H, Rao YN, Calisher CH (1992) Isolation of Getah virus from mosquitos collected on Hainan Island, China, and results of a serosurvey. Southeast Asian J Trop Med Public Health 23(4):730–734
Seo HJ, Kim HC, Klein TA, Park JY, Cho YS, Cho IS, Kyung SG, Yeh JY (2012) Characterization of recent Getah virus isolates from South Korea. Acta Virol 56(3):265–267
Turell MJ, O’Guinn ML, Wasieloski LP Jr, Dohm DJ, Lee WJ, Cho HW, Kim HC, Burkett DA, Mores CN, Coleman RE, Klein TA (2003) Isolation of Japanese encephalitis and Getah viruses from mosquitoes (Diptera: Culicidae) collected near Camp Greaves, Gyonggi Province, Republic of Korea, 2000. J Med Entomol 40(4):580–584
Zhai YG, Wang HY, Sun XH, Fu SH, Wang HQ, Attoui H, Tang Q, Liang GD (2008) Complete sequence characterization of isolates of Getah virus (genus Alphavirus, family Togaviridae) from China. J Gen Virol 89(Pt 6):1446–1456. https://doi.org/10.1099/vir.0.83607-0
Li YY, Fu SH, Guo XF, Lei WW, Li XL, Song JD, Cao L, Gao XY, Lyu Z, He Y, Wang HY, Ren XJ, Zhou HN, Wang GQ, Liang GD (2017) Identification of a Newly Isolated Getah Virus in the China-Laos Border, China. Biomed Environ Sci: BES 30(3):210–214. https://doi.org/10.3967/bes2017.028
Yago K, Hagiwara S, Kawamura H, Narita M (1987) A fatal case in newborn piglets with Getah virus infection: isolation of the virus. Nihon juigaku zasshi Jpn J Vet Sci 49(6):989–994
Shirako Y, Yamaguchi Y (2000) Genome structure of Sagiyama virus and its relatedness to other alphaviruses. J Gen Virol 81(Pt 5):1353–1360. https://doi.org/10.1099/0022-1317-81-5-1353
Shibata I, Hatano Y, Nishimura M, Suzuki G, Inaba Y (1991) Isolation of Getah virus from dead fetuses extracted from a naturally infected sow in Japan. Vet Microbiol 27(3–4):385–391
Wekesa SN, Inoshima Y, Murakami K, Sentsui H (2001) Genomic analysis of some Japanese isolates of Getah virus. Vet Microbiol 83(2):137–146
Romeiro MF, de Souza WM, Tolardo AL, Vieira LC, Henriques DA, de Araujo J, Siqueira CE, Colombo TE, Aquino VH, da Fonseca BA, de Morais Bronzoni RV, Nogueira ML, Durigon EL, Figueiredo LT (2016) A real-time RT-PCR for rapid detection and quantification of mosquito-borne alphaviruses. Arch Virol 161(11):3171–3177. https://doi.org/10.1007/s00705-016-3019-0
Kumanomido T, Wada R, Kanemaru T, Kamada M, Hirasawa K, Akiyama Y (1988) Clinical and virological observations on swine experimentally infected with Getah virus. Vet Microbiol 16(3):295–301
Sugiyama I, Shimizu E, Nogami S, Suzuki K, Miura Y, Sentsui H (2009) Serological survey of arthropod-borne viruses among wild boars in Japan. J Vet Med Sci 71(8):1059–1061
Acknowledgements
This work was supported by the National Key Research and Development Program of China (2017YFD0500104), the Technology and Development Program of Jilin Province (20150520127JH), Central Public-Interest Scientific Institution Basal Research Fund (16103420161034), and Key Laboratory of Preventive Veterinary Medicine of Education Department in Guangdong Province of China (2014KTSPT037).
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Shi, N., Liu, H., Li, Lx. et al. Development of a TaqMan probe-based quantitative reverse transcription PCR assay for detection of Getah virus RNA. Arch Virol 163, 2877–2881 (2018). https://doi.org/10.1007/s00705-018-3927-2
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DOI: https://doi.org/10.1007/s00705-018-3927-2