A polyprobe for the simultaneous detection by non-isotopic molecular hybridisation has been developed to detect any of the following six viruses causing important economic losses in tomato crops: Tomato spotted wilt virus, Tomato mosaic virus, Pepino mosaic virus, Cucumber mosaic virus, Potato Y virus and Parietaria mottle virus. The polyprobe detected all six viruses with similar sensitivity to that obtained using individual riboprobes. In addition, we evaluated the possible use of the tissue-printing as a sample preparation technique applied to routine diagnosis of tomato plants with the polyprobe.