Comparison of reverse transcription polymerase chain reaction, enzyme linked immunosorbent assay and virus isolation for the routine diagnosis of foot-and-mouth disease

• Published in: Journal of virological methods 1998-02, Vol.70 (2), p.213-217
• Main Authors: Reid, S.M, Forsyth, M.A, Hutchings, G.H, Ferris, N.P
• Format: Article
• Language: English
• Subjects: Animal viral diseases; Animals; ANTIGENE; ANTIGENOS; ANTIGENS; APHTHOVIRUS; BETAIL; Biological and medical sciences; Cattle; DIAGNOSIS; DIAGNOSTIC; DIAGNOSTICO; ELISA; Enzyme-Linked Immunosorbent Assay - methods; Enzyme-Linked Immunosorbent Assay - veterinary; FIEBRE AFTOSA; FIEVRE APHTEUSE; FOOT AND MOUTH DISEASE; Foot-and-Mouth Disease - diagnosis; Foot-and-Mouth Disease - virology; Fundamental and applied biological sciences. Psychology; GANADO; Goats; Infectious diseases; LIVESTOCK; Medical sciences; Microbiology; Milk - virology; PCR; Picornaviridae - isolation & purification; Polymerase chain reaction; Polymerase Chain Reaction - methods; Polymerase Chain Reaction - veterinary; Sheep; Swine; Techniques used in virology; TEST ELISA; Viral diseases; Virology; VIRUS FIEBRE AFTOSA; VIRUS FIEVRE APHTEUSE
• ISSN: 0166-0934; 1879-0984
• DOI: 10.1016/S0166-0934(97)00181-X

A reverse transcription polymerase chain reaction (RT–PCR) method was compared with virus isolation in cell culture and the antigen detection ELISA for the primary diagnosis of foot-and-mouth disease (FMD) on 166 clinical samples from the field. Eighty samples were positive by virus isolation/ELISA and 78 by RT–PCR. The RT–PCR detected FMD viral RNA in 11 of the 86 samples assessed as negative by virus isolation/ELISA but conversely failed to diagnose 13 samples identified as positive by the latter procedures. This RT–PCR is not serotype-specific so a cDNA product is indicative of the presence of FMD viral RNA only. Confirmation of the specificity of the cDNA product and the identification of the serotype requires nucleotide sequence analysis. The value of the RT–PCR is that it can rapidly facilitate the molecular analysis of field isolates and thus provide important epidemiological information regarding the source of outbreaks. However, it is a sophisticated technique requiring specialised equipment, expertise and refined reagents and has to be used in conjunction with current procedures for FMD diagnosis.