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Journal of Clinical Microbiology, December 2000, p. 4604-4613, Vol. 38, No. 12
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Development of Reverse Transcription-PCR (Oligonucleotide Probing) Enzyme-Linked Immunosorbent Assays for Diagnosis and Preliminary Typing of Foot-and-Mouth Disease: a New System Using Simple and Aqueous-Phase Hybridization

Soren Alexandersen,* Morag A. Forsyth, Scott M. Reid, and Graham J. Belsham

Institute for Animal Health, Pirbright Laboratory, Pirbright, Woking, Surrey, GU24 ONF, United Kingdom

Received 21 July 2000/Returned for modification 4 September 2000/Accepted 24 September 2000

A reverse transcription-PCR (RT-PCR)-enzyme-linked immunosorbent assay system that detects a relatively conserved region within the RNA genome of all seven serotypes of foot-and-mouth disease virus (FMDV) has been developed. The high specificity of the assay is achieved by including a rapid hybridization step with a biotin-labeled internal oligonucleotide. The assay is highly sensitive, fast, and easy to perform. A similar assay, based on a highly variable region of the FMDV genome and employing a single asymmetric RT-PCR and multiple hybridization oligonucleotides, was developed to demonstrate the method's ability to type FMDV. Based on our theoretical and practical knowledge of the methodology, we predict that similar assays are applicable to diagnosis and strain differentiation in any system amenable to PCR amplification.

* Corresponding author. Mailing address: Institute for Animal Health, Pirbright Laboratory, Pirbright, Woking, Surrey, GU24 ONF, United Kingdom. Phone: 44 1483 232 441. Fax: 44 1483 232 448. E-mail: soren.alexandersen{at}bbsrc.ac.uk.

Journal of Clinical Microbiology, December 2000, p. 4604-4613, Vol. 38, No. 12
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Wang, E., Paessler, S., Aguilar, P. V., Carrara, A.-S., Ni, H., Greene, I. P., Weaver, S. C. (2006). Reverse Transcription-PCR-Enzyme-Linked Immunosorbent Assay for Rapid Detection and Differentiation of Alphavirus Infections. J. Clin. Microbiol. 44: 4000-4008 [Abstract] [Full Text]  
  • Alexandersen, S., Oleksiewicz, M. B., Donaldson, A. I. (2001). The early pathogenesis of foot-and-mouth disease in pigs infected by contact: a quantitative time-course study using TaqMan RT-PCR. J. Gen. Virol. 82: 747-755 [Abstract] [Full Text]  


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