Diagnosis of foot-and mouth disease by real time reverse transcription polymerase cha

Background: Foot-and-mouth disease (FMD) is an economically important
and highly contagious viral disease that affects cloven-hoofed domestic
and wild animals. Virus isolation and enzyme-linked immunosorbent assay
(ELISA) are the gold standard tests for diagnosis of FMD. As these
methods are time consuming, assays based on viral nucleic acid
amplification have been developed. Results: A previously described
real-time reverse transcriptase polymerase chain reaction (RT-PCR)
assay with high sensitivity and specificity under laboratorial and
experimental conditions was used in the current study. To verify the
applicability of this assay under field conditions in Brazil, 460 oral
swabs from cattle were collected in areas free of FMD (n = 200) and
from areas with outbreaks of FMD (n = 260). Three samples from areas
with outbreaks of FMD were positive by real-time RT-PCR, and 2 of those
samples were positive by virus isolation and ELISA. Four other samples
were considered inconclusive by real-time RT-PCR (threshold cycle [Ct]
> 40); whereas all 200 samples from an area free of FMD were
real-time RT-PCR negative. Conclusion: real-time RT-PCR is a powerful
technique for reliable detection of FMDV in a fraction of the time
required for virus isolation and ELISA. However, it is noteworthy that
lack of infrastructure in certain areas with high risk of FMD may be a
limiting factor for using real-time RT-PCR as a routine diagnostic tool.

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