This study utilized the real‐time PCR primers, probes, master mixes and cycling conditions that have been previously described (Bowden et al., 2008). Pilot studies were undertaken to assess the performance of newly developed lyophilized PCR reagents that were prepared and assembled into assay cartridges by Enigma Diagnostics (Salisbury, UK). A decimal dilution series (Neat to 10−10) of DNA prepared from an LSDV isolate (Israel LSD‐07 POX‐V1‐07‐08, isolated from naturally infected cattle in 2007) was prepared in nuclease‐free water containing carrier RNA (1 μg ml−1). In these experiments, 5 μl of each dilution was mixed with 20 μl of nuclease‐free water and this was used to re‐suspend the lyophilized reagent pellets prepared by Enigma Diagnostics. This 25 μl suspension was then transferred into a 96‐well PCR plate. For the conventional wet reagents, 2 μl of DNA was added to 18 μl of diagnostic assay mastermix (Bowden et al., 2008) prior to transfer into 96‐well PCR plate. This initial laboratory validation of lyophilized reagents was carried out on the Mx3005P quantitative PCR machine (Stratagene). Parallel testing demonstrated an improved analytical sensitivity of one log10 for the lyophilized reagents when compared to the reference test (Fig. 1a). This one log10 increase in analytical sensitivity was maintained when the new lyophilized assay was applied to a decimal dilution series (10−1–10−10) of Israel LSD‐07 POX‐V1‐07‐08 virus prepared in 10% w/v homogenized cattle skin suspensions and run in full, including DNA extraction on the Enigma FL (Fig. 1b). For the above comparison, one aliquot per dilution was extracted on the MagNA Pure LC Robot (Roche) using the Total Nucleic Acid Isolation kit (Roche) following manufacturer guidelines, followed by real‐time PCR performed using wet reagents assayed on the Mx3005P quantitative PCR machine (Stratagene) (reference test). The extraction and real‐time PCR for the second aliquot (0.5 ml) was performed in a complete automated cycle on the Enigma FL.