On the reliability of recombinase polymerase amplification - lateral flow assay using ITS rRNA gene primers and probe as a new detection method of the golden potato cyst nematode, Globodera rostochiensis

. Recombinase polymerase amplification (RPA) is an isothermal in vitro nucleic acid amplification technique, which has been adopted as a novel molecular technology for simple, rapid, reliable, and low-resource diagnostics of nematodes and other damaging organisms. Recently, RPA integrated with lateral flow assay (LFA) using the ITS rRNA gene primers and probe was proposed by Wang et al. (2022) as a new detection method of Globodera rostochiensis. The reliability of any diagnostic method should be validated on a broad range of populations of different species from different geographical locations as well as on related taxa to avoid erroneous identification with false positive results. A total of 24 isolates of 13 cyst nematode species were used to validate the analytical specificity of the method mentioned above. The results of these tests showed that the ITS rRNA gene primers and probe did not give specific RPA-LFA detection for the golden potato cyst nematode and gave positive reactions not only with G. rostochiensis , but also with other Globodera parasitising solanaceous plants, G. artemisiae , and representatives of the genus Punctodera parasitising grasses.