RT-PCR in foot-and-mouth disease diagnosis.
A RT-PCR assay for the specific detection of RNA sequences from foot-and-mouth disease virus (FMDV) has been developed. The procedure permits also the detection of sequences that correlate with established FMDV serotypes. A computer program that allows selection of genotype-specific primers for RT-PCR amplification was used for the identification of FMDV specific sequences for PCR amplification on RNA replicase (3D) gene regions. Specific, rapid and highly sensitive detection was achieved for a wide collection of RNA samples from FMDV types C, A and O, either purified from tissue culture or extracted from lesions of infected animals. Similarly, serotype-specific primers were designed to amplify the carboxy-terminal end of the VP1 gene of FMDV types either C, A or O. The results of PCR amplification of different FMDV RNAs using type-specific primers are in agreement with the serological typing of the corresponding viruses. A combination of this approach with a simplified sample processing, carried out following direct adsorption of viral suspensions to microtiter plates, provides a rapid, reliable method of viral diagnosis.
Núñez
J I
JI
Centro de Investigación en Sanidad Animal, INIA, Madrid, Spain.
Blanco
E
E
Hernández
T
T
Dopazo
J
J
Sobrino
F
F
D000818:Animals; D005537:Aphthovirus / Q000235:genetics*; D005536:Foot-and-Mouth Disease / Q000175:diagnosis* / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*; D012367:RNA, Viral / Q000032:analysis / Q000235:genetics*; D012324:RNA-Dependent RNA Polymerase / Q000235:genetics; D012680:Sensitivity and Specificity; D017423:Sequence Analysis, RNA; D013552:Swine
D012367:RNA, Viral; D012324:RNA-Dependent RNA Polymerase