Direct PCR detection of foot-and-mouth disease virus.
A PCR assay for the detection and characterization of foot-and-mouth disease virus was developed. The procedure allows RT-PCR amplification following direct adsorption of viral suspensions to microtiter plates, avoiding previous steps of phenol-extraction or heating. Using this procedure, FMDV-specific (based on 3D gene sequences), as well as serotype-specific (based on VP1 gene sequences) amplification were achieved for viral samples of serotypes A, O and C, either from cell culture supernatants or from lesions of infected animals. The assay allowed detection of around 15 PFU, being 500-fold more sensitive than a conventional indirect ELISA. This new method constitutes a simple, rapid and efficient alternative for the diagnosis and characterization of FMDV by PCR.
Rodríguez
A
A
Centro de Investigación en Sanidad Animal, INIA, Valdeolmos, Madrid, Spain.
Núñez
J I
JI
Nolasco
G
G
Ponz
F
F
Sobrino
F
F
de Blas
C
C
D000818:Animals; D005537:Aphthovirus / Q000235:genetics / Q000302:isolation & purification*; D005536:Foot-and-Mouth Disease / Q000175:diagnosis*; D016133:Polymerase Chain Reaction / Q000379:methods