The greatest problem facing the diagnostic application of PCR is the production of false positive results. They are attributable to contamination by nucleic acids, particularly from the previously amplified material (carry over). Any contaminant, even the smallest airborne remnant carried over from the previous PCR procedure or from a strongly positive sample (contamination), may be multiplied and produce a false positive results. In the Real-time PCR, the problem of carry-over is significantly reduced because of the real-time measuring principle, which is based on closed tube system. In order to increase assay throughput and minimise operator errors, real-time PCR assays for FMDV have also been automated using robots for nucleic acid extraction and liquid handling equipment to setup the reaction mixes [146, 169] . The assay has also been applied for FMDV and it is capable of detecting all seven serotypes [39, 161, [169] [170] [171] [172] [173] [174] [175] [176] [177] . First interassay/laboratory equivalence investigation [178] , participated by five European reference laboratories for detection of FMDV has observed that the best of the Real-Time PCR assays used in each laboratory gave comparable results unlike the VI results which were highly variable. Performance of three real-time instruments: the LightCycler.2 (Roche), the SmartCycler I I (Cepheid), and the SDS 7900HT (AB) was compared for detection of FMDV and has successfully identified the FMDV genome and beta actin mRNA from several sources of infected nasal and oral swabs, as well as probang samples [167] . In addition to extensive testing of vesicular epithelium samples, the performance of rRT-PCR has recently been assessed on milk [179] and "probang" samples [180] . Automated real-time RT-PCR has been compared with VI and antigen-detection ELISA with result being more positive by real-time PCR indicating that the real-time PCR has higher sensitivity than VI for the detection of FMDV in epithelial samples [181] .