Phytopathogenic Mollicutes

Real-time quantitative PCR is the most common technique for detection of phytoplasma, small bacteria associated with several economically important plant diseases worldwide. Infection cycles of phytoplasma-associated diseases involve multitrophic interactions between the bacterium and different hosts, i.e. insect vectors and plants, thus diagnostic analyses must be done in different DNA matrices. For routine diagnostics of plant or insect samples by qPCR it is crucial to have an endogenous internal control for the host DNA. While several host specific primer and probe combinations are available as endogenous control for phytoplasma detection, a universal internal control for different eukaryotic host organisms is lacking. Thus a universal internal control that can be used in a broad range of phytoplasma host species was developed and the use of defined strict criteria for the evaluation of qPCR results are elucitated.