This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Wakeley, P. R.
Right arrow Articles by Fooks, A. R.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Wakeley, P. R.
Right arrow Articles by Fooks, A. R.

 Previous Article  |  Next Article 

Journal of Clinical Microbiology, June 2005, p. 2786-2792, Vol. 43, No. 6
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.6.2786-2792.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Development of a Real-Time, TaqMan Reverse Transcription-PCR Assay for Detection and Differentiation of Lyssavirus Genotypes 1, 5, and 6

P. R. Wakeley,* N. Johnson, L. M. McElhinney, D. Marston, J. Sawyer, and A. R. Fooks

Rabies Research and Diagnostic Group, World Health Organization Collaborating Centre for the Characterization of Rabies and Rabies-Related Viruses, Department of Virology, and Technology Transfer Unit, Biotechnology Department, Veterinary Laboratories Agency, New Haw, Addlestone, Surrey KT15 3NB, United Kingdom

Received 21 July 2004/ Returned for modification 23 November 2004/ Accepted 17 January 2005

Several reverse transcription-PCR (RT-PCR) methods have been reported for the detection of rabies and rabies-related viruses. These methods invariably involve multiple transfers of nucleic acids between different tubes, with the risk of contamination leading to the production of false-positive results. Here we describe a single, closed-tube, nonnested RT-PCR with TaqMan technology that distinguishes between classical rabies virus (genotype 1) and European bat lyssaviruses 1 and 2 (genotypes 5 and 6) in real time. The TaqMan assay is rapid, sensitive, and specific and allows for the genotyping of unknown isolates concomitant with the RT-PCR. The assay can be applied quantitatively and the use of an internal control enables the quality of the isolated template to be assessed. Despite sequence heterogeneity in the N gene between the different genotypes, a universal forward and reverse primer set has been designed, allowing for the simplification of previously described assays. We propose that within a geographically constrained area, this assay will be a useful tool for the detection and differentiation of members of the Lyssavirus genus.

* Corresponding author. Mailing address: Technology Transfer Unit, Veterinary Laboratories Agency-Weybridge, New Haw, Addlestone, Surrey KT15 3NB, United Kingdom. Phone: 44 (0)1932 357759. Fax: 44 (0)1932 357890. E-mail: p.wakeley{at}vla.defra.gsi.gov.uk.

Journal of Clinical Microbiology, June 2005, p. 2786-2792, Vol. 43, No. 6
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.6.2786-2792.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Freuling, C., Vos, A., Johnson, N., Kaipf, I., Denzinger, A., Neubert, L., Mansfield, K., Hicks, D., Nunez, A., Tordo, N., Rupprecht, C. E., Fooks, A. R., Muller, T. (2009). Experimental infection of serotine bats (Eptesicus serotinus) with European bat lyssavirus type 1a. J. Gen. Virol. 90: 2493-2502 [Abstract] [Full Text]  
  • Horton, D. L., Voller, K., Haxton, B., Johnson, N., Leech, S., Goddard, T., Wilson, C., McElhinney, L. M., Fooks, A. R. (2009). European bat lyssavirus type 2 in a Daubenton's bat in Scotland. Vet Rec. 165: 383-384 [Full Text]  
  • Johnson, N., Vos, A., Neubert, L., Freuling, C., Mansfield, K. L., Kaipf, I., Denzinger, A., Hicks, D., Nunez, A., Franka, R., Rupprecht, C. E., Muller, T., Fooks, A. R. (2008). Experimental study of European bat lyssavirus type-2 infection in Daubenton's bats (Myotis daubentonii). J. Gen. Virol. 89: 2662-2672 [Abstract] [Full Text]  
  • Franka, R., Johnson, N., Muller, T., Vos, A., Neubert, L., Freuling, C., Rupprecht, C. E., Fooks, A. R. (2008). Susceptibility of North American big brown bats (Eptesicus fuscus) to infection with European bat lyssavirus type 1. J. Gen. Virol. 89: 1998-2010 [Abstract] [Full Text]  
  • Fooks, A. R., Harkess, G., Goddard, T., Marston, D. A., McElhinney, L., Brown, K., Morgan, D., Paul, R., Thomas, P. J., Smith, B. (2008). Rabies virus in a dog imported to the UK from Sri Lanka. Vet Rec. 162: 598-598 [Full Text]  
  • Harris, S. L., Mansfield, K., Marston, D. A., Johnson, N., Pajamo, K., O'Brien, N., Black, C., McElhinney, L. M., Fooks, A. R. (2007). Isolation of European bat lyssavirus type 2 from a Daubenton's bat (Myotis daubentonii) in Shropshire. Vet Rec. 161: 384-386 [Full Text]  
  • Fooks, A. R., Marston, D., Parsons, G., Earl, D., Dicker, A., Brookes, S. M. (2006). Isolation of EBLV-2 in a Daubenton's bat (Myotis daubentonii) found in Oxfordshire. Vet Rec. 159: 534-535 [Full Text]  
  • Solomon, T., Marston, D., Mallewa, M., Felton, T., Shaw, S., McElhinney, L. M, Das, K., Mansfield, K., Wainwright, J., Kwong, G. N. M., Fooks, A. R (2005). Paralytic rabies after a two week holiday in India. BMJ 331: 501-503 [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»