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

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 asymmetry: RT-PCR and multiple hybridization oligonucleotides, was developed to demonstrate the method's ability to type FMDV. Based an our theoretical and practical knowledge of the methadology, we predict that similar assays are applicable to diagnosis and strain differentiation in any system amenable to PCR amplification.