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 acid aqueous-phase hybridization

A reverse transcription-PCR (RT-PCR)-enzyme-linked immunosorbent assay syst em 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 hybridiz ation step with a biotin-labeled internal oligonucleotide. The assay is hig hly sensitive, fast, and easy to perform, A similar assay, based on a highl y variable region of the FMDV genome and employing a single asymmetry: RT-P CR and multiple hybridization oligonucleotides, was developed to demonstrat e the method's ability to type FMDV. Based an our theoretical and practical knowledge of the methadology, we predict that similar assays are applicabl e to diagnosis and strain differentiation in any system amenable to PCR amp lification.