Development of a one‐step multiplex reverse transcription‐polymerase chain reaction and Luminex xTAG assay for the simultaneous detection of yellowing viruses infecting sugar beet

Yellowing viruses are an increasing threat to sugar beet cultivation, due to limitations on insecticide usage and climate change. Virus detection, monitoring and resistance breeding are key to secure high sugar beet yields in the future. For this research, a one‐step multiplex reverse transcription (mRT)‐PCR method was designed to detect simultaneously beet mild yellowing virus, beet chlorosis virus, beet yellows virus (BYV), beet mosaic virus and turnip yellows virus. The addition of Luminex xTAG array technology was used as a follow‐up method to increase assay specificity. The one‐step mRT‐PCR was evaluated on 22 field samples with single and mixed virus infections. The xTAG assay works as expected both in a simplex and multiplex setting, except that BYV detection needs optimization in the multiplex setting. In the future, the Luminex xTAG assay would be an excellent method for the detection of beet yellowing viruses due to its high specificity and the potential to increase the number of targets.