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Evaluation of the portable Cepheid SmartCycler real‐time PCR machine for the rapid diagnosis of foot‐and‐mouth disease
Author(s) -
Hearps A.,
Zhang Z.,
Alexandersen S.
Publication year - 2002
Publication title -
veterinary record
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.261
H-Index - 99
eISSN - 2042-7670
pISSN - 0042-4900
DOI - 10.1136/vr.150.20.625
Subject(s) - taqman , foot and mouth disease virus , real time polymerase chain reaction , virology , foot and mouth disease , chromatography , biology , medicine , virus , chemistry , biochemistry , gene
The ability of the portable Cepheid SmartCycler real‐time PCR machine to detect foot‐and‐mouth disease (FMD) virus sensitively and accurately was evaluated by comparing the results of the analyses of nasal swab and serum samples from experimentally infected animals with those obtained from the real‐time PCR assay currently in use in the laboratory. The results indicated that the ability of the machine to detect viral RNA iS greatly affected by the PCR reagents used for the assay. When it was used with PCR beads it was unable to detect weakly positive samples, but when TaqMan core reagents were used for the assay, its sensitivity was significantly increased. The machine could be used for the laboratory‐based detection of FMD; however, as with all assays, significant optimisation of assay conditions as well as solid validation of the technique is required.