Construction of a reverse genetic system for porcine astrovirus

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Abstract

In order to construct a full-length infectious cDNA clone of porcine astrovirus, three fragments covering the complete genome of PAstV1-GX1 strain were amplified by RT-PCR. All three PCR-amplified fragments were cloned into T-Vector pMD19 (Simple), and subsequently assembled into a full-length cDNA clone by subcloning. A silent nucleotide change creating a PstI site was engineered into the full-length cDNA clone to distinguish the rescued virus from the parental virus. Upon transfection of BHK-21 cells with the in vitro transcripts of both the original and constructed cDNAs, typical cytopathic effects were observed on PK-15 cells after serial passaging of the cell supernatant. The construction and recovery of the infectious cDNA clone of porcine astrovirus will provide a valuable experimental system to study the genome function and pathogenesis of astroviruses.

Notes

Acknowledgements

We thank Yizhi Zhong for suggestions on the infectious clone construction. We are grateful to many colleagues for technical assistance and to Dr. Dev Sooranna, Imperial College London, for editing the manuscript. This work was supported by the National Natural Science Foundation of China (No. 31460671 and 31760735) and Innovation Driven Development Special Foundation of Guangxi (AA17204057-1).

Compliance with ethical standards

Conflict of interest

The authors declare they have no conflict of interest.

References

  1. 1.
    Alfred N, Liu H, Li ML, Hong SF, Tang HB, Wei ZZ, Chen Y, Li FK, Zhong YZ, Huang WJ (2015) Molecular epidemiology and phylogenetic analysis of diverse bovine astroviruses associated with diarrhea in cattle and water buffalo calves in China. J Vet Med Sci 77:643–651CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Arruda B, Arruda P, Hensch M, Chen Q, Zheng Y, Yang C, Gatto IRH, Ferreyra FM, Gauger P, Schwartz K, Bradner L, Harmon K, Hause B, Li G (2017) Porcine astrovirus type 3 in central nervous system of swine with polioencephalomyelitis. Emerg Infect Dis 23:2097–2100CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Bass DM, Qiu S (2000) Proteolytic processing of the astrovirus capsid. J Virol 74:1810–1814CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Blomström A-L, Ley C, Jacobson M (2014) Astrovirus as a possible cause of congenital tremor type AII in piglets? Acta Vet Scand 56:82CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Bridger J (1980) Detection by electron microscopy of caliciviruses, astroviruses and rotavirus-like particles in the faeces of piglets with diarrhoea. Vet Rec 107:532–533PubMedGoogle Scholar
  6. 6.
    Brnic D, Prpic J, Keros T, Roic B, Staresina V, Jemersic L (2013) Porcine astrovirus viremia and high genetic variability in pigs on large holdings in Croatia. Infect Genet Evol 14:258–264CrossRefPubMedGoogle Scholar
  7. 7.
    Brnic D, Jemersic L, Keros T, Prpic J (2014) High prevalence and genetic heterogeneity of porcine astroviruses in domestic pigs. Vet J 202:390–392CrossRefPubMedGoogle Scholar
  8. 8.
    Brown JR, Morfopoulou S, Hubb J, Emmett WA, Ip W, Shah D, Brooks T, Paine SM, Anderson G, Virasami A, Tong CY, Clark DA, Plagnol V, Jacques TS, Qasim W, Hubank M, Breuer J (2015) Astrovirus VA1/HMO-C: an increasingly recognized neurotropic pathogen in immunocompromised patients. Clin Infect Dis 60:881–888CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Cai Y, Yin W, Zhou Y, Li B, Ai L, Pan M, Guo W (2016) Molecular detection of Porcine astrovirus in Sichuan Province. China. Virol J 13:6CrossRefPubMedGoogle Scholar
  10. 10.
    De Benedictis P, Schultz-Cherry S, Burnham A, Cattoli G (2011) Astrovirus infections in humans and animals - molecular biology, genetic diversity, and interspecies transmissions. Infect Genet Evol 11:1529–1544CrossRefPubMedGoogle Scholar
  11. 11.
    Finkbeiner SR, Holtz LR, Jiang Y, Rajendran P, Franz CJ, Zhao G, Kang G, Wang D (2009) Human stool contains a previously unrecognized diversity of novel astroviruses. Virol J 6:161CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Finkbeiner SR, Li Y, Ruone S, Conrardy C, Gregoricus N, Toney D, Virgin HW, Anderson LJ, Vinje J, Wang D, Tong S (2009) Identification of a novel astrovirus (astrovirus VA1) associated with an outbreak of acute gastroenteritis. J Virol 83:10836–10839CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Geigenmuller U, Ginzton NH, Matsui SM (1997) Construction of a genome-length cDNA clone for human astrovirus serotype 1 and synthesis of infectious RNA transcripts. J Virol 71:1713–1717PubMedPubMedCentralGoogle Scholar
  14. 14.
    Karlsson EA, Small CT, Freiden P, Feeroz MM, Matsen FAT, San S, Hasan MK, Wang D, Jones-Engel L, Schultz-Cherry S (2015) Non-human primates harbor diverse mammalian and avian astroviruses including those associated with human infections. PLoS Pathog 11:e1005225CrossRefPubMedPubMedCentralGoogle Scholar
  15. 15.
    Lan D, Ji W, Shan T, Cui L, Yang Z, Yuan C, Hua X (2011) Molecular characterization of a porcine astrovirus strain in China. Arch Virol 156:1869–1875CrossRefPubMedGoogle Scholar
  16. 16.
    Liu N, Wang F, Shi J, Zheng L, Wang X, Zhang D (2014) Molecular characterization of a duck hepatitis virus 3-like astrovirus. Vet Microbiol 170:39–47CrossRefPubMedGoogle Scholar
  17. 17.
    Luo Z, Roi S, Dastor M, Gallice E, Laurin M, L’homme Y (2011) Multiple novel and prevalent astroviruses in pigs. Vet Microbiol 149:316–323CrossRefPubMedGoogle Scholar
  18. 18.
    Méndez E, Murillo A, Velázquez R, Burnham A, Arias CF (2013) Replication Cycle of Astroviruses. In: Schultz-Cherry S (ed) Astrovirus research: essential ideas, everyday impacts, future directions. Springer, New York, pp 19–45Google Scholar
  19. 19.
    Madeley CR, Cosgrove BP (1975) Letter: 28 nm particles in faeces in infantile gastroenteritis. Lancet 2:451–452CrossRefPubMedGoogle Scholar
  20. 20.
    Meliopoulos VA, Kayali G, Burnham A, Oshansky CM, Thomas PG, Gray GC, Beck MA, Schultz-Cherry S (2014) Detection of antibodies against Turkey astrovirus in humans. PLoS One 9:e96934CrossRefPubMedPubMedCentralGoogle Scholar
  21. 21.
    Meulenberg JJ, Bos-de Ruijter JN, van de Graaf R, Wensvoort G, Moormann RJ (1998) Infectious transcripts from cloned genome-length cDNA of porcine reproductive and respiratory syndrome virus. J Virol 72:380–387PubMedPubMedCentralGoogle Scholar
  22. 22.
    Mor SK, Chander Y, Marthaler D, Patnayak DP, Goyal SM (2012) Detection and molecular characterization of Porcine astrovirus strains associated with swine diarrhea. J Vet Diagn Invest 24:1064–1067CrossRefPubMedGoogle Scholar
  23. 23.
    Naccache SN, Peggs KS, Mattes FM, Phadke R, Garson JA, Grant P, Samayoa E, Federman S, Miller S, Lunn MP, Gant V, Chiu CY (2015) Diagnosis of neuroinvasive astrovirus infection in an immunocompromised adult with encephalitis by unbiased next-generation sequencing. Clin Infect Dis 60:919–923CrossRefPubMedPubMedCentralGoogle Scholar
  24. 24.
    Oem J-K, An D-J (2014) Phylogenetic analysis of bovine astrovirus in Korean cattle. Virus Genes 48:372–375CrossRefPubMedGoogle Scholar
  25. 25.
    Padmanabhan A, Hause BM (2016) Detection and characterization of a novel genotype of porcine astrovirus 4 from nasal swabs from pigs with acute respiratory disease. Arch Virol 161:2575–2579CrossRefPubMedGoogle Scholar
  26. 26.
    Patel AK, Pandit RJ, Thakkar JR, Hinsu AT, Pandey VC, Pal JK, Prajapati KS, Jakhesara SJ, Joshi CG (2017) Complete genome sequence analysis of chicken astrovirus isolate from India. Vet Res Commun 41:67–75CrossRefPubMedGoogle Scholar
  27. 27.
    Pfaff F, Schlottau K, Scholes S, Courtenay A, Hoffmann B, Hoper D, Beer M (2017) A novel astrovirus associated with encephalitis and ganglionitis in domestic sheep. Transbound Emerg Dis 64:677–682CrossRefPubMedGoogle Scholar
  28. 28.
    Rivera R, Nollens HH, Venn-Watson S, Gulland FM, Wellehan JF Jr (2010) Characterization of phylogenetically diverse astroviruses of marine mammals. J Gen Virol 91:166–173CrossRefPubMedGoogle Scholar
  29. 29.
    Schlottau K, Schulze C, Bilk S, Hanke D, Hoper D, Beer M, Hoffmann B (2016) Detection of a Novel Bovine Astrovirus in a Cow with Encephalitis. Transbound Emerg Dis 63:253–259CrossRefPubMedGoogle Scholar
  30. 30.
    Seuberlich T, Wuthrich D, Selimovic-Hamza S, Drogemuller C, Oevermann A, Bruggmann R, Bouzalas I (2016) Identification of a second encephalitis-associated astrovirus in cattle. Emerg Microbes Infect 5:e5CrossRefPubMedCentralGoogle Scholar
  31. 31.
    Singh A, Mor SK, Jindal N, Patnayak D, Sobhy NM, Luong NT, Goyal SM (2016) Detection and molecular characterization of astroviruses in turkeys. Adv Virol 161:939–946Google Scholar
  32. 32.
    Sun N, Yang Y, Wang G-S, Shao X-Q, Zhang S-Q, Wang F-X, Tan B, Tian F-L, Cheng S-P, Wen Y-J (2014) Detection and characterization of avastrovirus associated with diarrhea isolated from minks in China. Food Environ Virol 6:169–174CrossRefPubMedGoogle Scholar
  33. 33.
    Takano T, Takashina M, Doki T, Hohdatsu T (2015) Detection of canine astrovirus in dogs with diarrhea in Japan. Arch Virol 160:1549–1553CrossRefPubMedGoogle Scholar
  34. 34.
    Thumfart JO, Meyers G (2002) Feline calicivirus: recovery of wild-type and recombinant viruses after transfection of cRNA or cDNA constructs. J Virol 76:6398–6407CrossRefPubMedPubMedCentralGoogle Scholar
  35. 35.
    Ulloa JC, Gutiérrez MF (2010) Genomic analysis of two ORF2 segments of new porcine astrovirus isolates and their close relationship with human astroviruses. Can J Microbiol 56:569–577CrossRefPubMedGoogle Scholar
  36. 36.
    Willcocks MM, Carter MJ (1993) Identification and sequence determination of the capsid protein gene of human astrovirus serotype 1. FEMS Microbiol Lett 114:1–7CrossRefPubMedGoogle Scholar
  37. 37.
    Xiao CT, Gimenez-Lirola LG, Gerber PF, Jiang YH, Halbur PG, Opriessnig T (2013) Identification and characterization of novel porcine astroviruses (PAstVs) with high prevalence and frequent co-infection of individual pigs with multiple PAstV types. J Gen Virol 94:570–582CrossRefPubMedGoogle Scholar
  38. 38.
    Xiao J, Li J, Hu G, Chen Z, Wu Y, Chen Y, Chen Z, Liao Y, Zhou J, Ke X, Ma L, Liu S, Zhou J, Dai Y, Chen H, Yu S, Chen Q (2011) Isolation and phylogenetic characterization of bat astroviruses in southern China. Arch Virol 156:1415–1423CrossRefPubMedGoogle Scholar
  39. 39.
    Yunus MA, Chung LM, Chaudhry Y, Bailey D, Goodfellow I (2010) Development of an optimized RNA-based murine norovirus reverse genetics system. J Virol Methods 169:112–118CrossRefPubMedPubMedCentralGoogle Scholar
  40. 40.
    Zaraket H, Abou-El-Hassan H, Kreidieh K, Soudani N, Ali Z, Hammadi M, Reslan L, Ghanem S, Hajar F, Inati A, Rajab M, Fakhouri H, Ghanem B, Baasiri G, Melhem NM, Dbaibo G (2017) Characterization of astrovirus-associated gastroenteritis in hospitalized children under five years of age. Infect Genet Evol 53:94–99CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag GmbH Austria, part of Springer Nature 2018

Authors and Affiliations

  1. 1.College of Animal Science and TechnologyGuangxi UniversityNanningPeople’s Republic of China

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