isrjournals
User Sign in    Editor Sign in

application of nucleic acid based techniques for detection of classical swine fever virus: a comparative study

ELINA KHATOON,N.N. BARMAN,MANAB DEKA,GITIKA RAJBONGSHI,KONGKON BARUAH,NIPU DEKA,D.P. BORA

Published in International Journal of Advanced Research in Computer Networking,Wireless and Mobile Communications

ISSN: 2320-7248          Impact Factor:1.8         Volume:2         Issue:2         Year: 01 April,2015         Pages:125-129

International Journal of Advanced Research in Computer Networking,Wireless and Mobile Communications

Abstract

Classical swine fever (CSF) is a contagious and devastating viral disease, causing serious losses in the pig industry worldwide. In the control programmes of CSF, rapid detection and identification of the causative agent is a crucial step. Various PCR based techniques like nested RT-PCR, SybrGreen based Real-Time PCR and TaqMan based Realtime PCR were used for detection of CSFV nucleic acid in clinical as well as tissue samples. In our study three detection systems were tested for classical swine fever virus (CSFV) detection and for its discrimination from other pestiviruses; Nested PCR, non-specific dsDNA-binding SYBR Green dye based and specific fluorogenic TaqMan MGB probes based Real-Time PCR. All three detection assays were evaluated for diagnosis of different field strains. However, one-step TaqMan Real-Time PCR assay was shown to be the most appropriate for pestivirus discrimination in comparison to the other two assays, moreover, it reduces the risk of contamination and is less time consuming.

Kewords

CSFV; SybrGreen dye; Nested PCR; TaqMan probe

Reference

[1]. B. Hoffmann, M. Beer: Validation of a real-time RT-PCR assay for sensitive and specific detection of classical swine fever. J Virol Methods, 2005, 130(1-2): pp. 36_44. [2]. P. Lowings, G. Ibata, J. Needham and D. Paton, D: Classical swine fever virus diversity and evolution. J.Gen. Virol., 1996, 77: pp. 1311_1321. [3]. G. Meyers, H.-J. Thiel, T. Rümenapf: Classical swine fever virus: recovery of infectious viruses from cDNA constructs and generation of recombinant cytopathogenic defective interfering particles. J. Virol., 1996, 70, pp.1588_1595. [4]. T. Rümenapf, H.-J. Thiel: Molecular biology of pestiviruses. In: Mettenleiter, T.C., Sobrino, F. (Eds.), Animal Viruses: Molecular Biology. CaisterAcademic Press, Norfolk, UK, 2008, pp. 39_96. [5]. G. Saikumar: The DBT Network project on Classical Swine fever with special reference to North Eastern Region, IVRI, Izatnagar (U.P.) [6]. H.-J. Thiel, M.S. Collett, E.A. Gould, F.X. Heinz, M. Houghton, G. Meyers, R.H. Purcell, C.M. Rice: Family flaviviridae. In: Fauquet, C.M., Mayo,M.A., Maniloff, J., Desselberger, U., Ball, L.A. (Eds.), Virus Taxonomy: VIIIth Report of the International Committee on Taxonomy of Viruses. Academic Press, San Diego, 2005, pp. 979_996. [7]. N.N. Barman, R.S. Gupt, N.K. Singh, A.K. Tiwari, R.K. Singh, S.K. Das: Comparative evaluation of molecular and antibody based technique for detection of classical swine fever virus infecting pigs of NE region, India. The Indian Journal of Animal Sciences, 2009; Vol 79, No. 10 [8]. D.J. Paton, A. McGoldrick, S. Belak, C. Mittelholzer, F. Koenen, H. Vanderhallen, M. Biagetti, G. De Mia, T. Stadejek, M.A. Hofmann, B. Thuer. Classical swine fever virus: a ring test to evaluate RT-PCR detection methods. Vet. Microbiol. 73, 2009b, pp. 159_174. [9] H.J. Thiel, R. Stark, E. Weiland, T. Rumenapf, G. Meyers: Hog cholera virus: Molecular composition of virions from a pestivirus. J Virol. 1991, 65(9), pp. 4705–4712