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Grapevine cultivation is endangered by several pathogens disseminated in latent form by propagating material. These pathogens include several viroids, viruses, phytoplasmas, bacteria and fungi (Bisztray et al. 2012, Martelli 2014). Their presence in the plant frequently cannot be recognized by symptoms in the plantations, but disease outbreak may take place in the later stage of infection or in the new plantations under favourable environmental conditions. Therefore the use of healthy planting material has a vital importance. Pathogen-free grapevine stocks can be selected from the existing plantations by various diagnostic protocols or produced by various curative (e. g., hot water) treatments and in vitro shoot tip and/or apical meristem cultures (Bisztray et al. 2012). The pathogen-free status of hot water treated and micropropagated plants should also be tested by appropriate diagnostic methods. During the recent decades the polymerase chain reaction (PCR) has become the most common diagnostic protocol in plant pathology. The introduction of the conventional PCR (including multiplex, nested and reverse trascription PCR) has opened wide application possibilities due to its simplicity and cost effectiveness. Later, it was followed by the more sensitive, but also more costly, quantitative realtime PCR (qRT-PCR). Quantitative real-time PCR protocols have already been developed for several viroids (Papayiannis 2014, Sun et al. 2014) , viruses (Harper et al. 2011, Osman & Rowhani 2008, Pacificio et al. 2011), phytoplasmas (Angelini 2010, Fahrentrapp et al. 2013, Pelletier et al. 2009), bacteria (Dreo et al. 2007, Harper et al. 2010, Johnson et al. 2013) and fungi (Martin et al. 2012, Schena et al. 2004). These protocols are based on the amplification of given nucleic acid sequences derived from plants or isolated microbes. Since viroids, viruses and phytoplasmas are closely associated with plant cells they are detected from total plant nucleic acid (RNA or DNA) preparations. Bacteria and fungi can be recovered from grapevine bleeding sap or from plant tissues on appropriate culture media thus they can be detected and identified both from total plant DNA and pure cultures. Recently, a medium for recovery of axenic phytoplasma cultures has also been developed (Contaldo et al. 2012) that will allow the identification of phytoplasmas from isolated colonies as well in the future. For nucleic acid (RNA or DNA) isolation several protocols have been developed during the last decades. For rapid field sampling the use of FTA cards (Whatman) can be proposed that allows a simple collection, transport and storage of nucleic acids derived from tissue sap or from homogenized tissues. Such samples then can be processed in PCR (Grund et al. 2010) to detect plant pathogens. Kits specifically developed for plant nucleic acids isolation (e. g., Spectrum Plant Total RNA Kit produced by SigmaAldrich and RNeasy Plant RNA Mini Kit produced by Qiagen) are also widely used for diagnostic purposes (Cseh 2012, Gambino & Gribaudo 2006, Li et al. 2011, Nassuth et Conventional PCR primers for the detection of grapevine pathogens disseminated by propagating material