Development of Multiplex RT-PCR Assay for the Simultaneous Detection of Four Systemic Diseases Infecting Citrus

The citrus systemic diseases, including Huanglongbing (caused by Candidatus Liberibacter asiaticus (CLas)), tristeza virus (CTV)), tatter leaf (CTLV)), and exocortis viroid (CEVd)), are threats to production in Taiwan. Reliable diagnostic methods important for the management of these diseases. In this study, we developed a multiplex reverse transcription–polymerase chain reaction (RT-PCR) assay detect four pathogens simultaneously. Herein, specific amplicons from each pathogen (295 bp CLas, 468 CTV, 120 CTLV, 196 CEVd) were successfully produced using optimized RT-PCR described here. sensitivity evaluation showed that low titers could be detected RT-PCR. Compared with published simplex assays, detection field samples study better performance. detections revealed commonly found fields, 30.0% mix-infected. To our knowledge, is first survey diseases Taiwan, it provides insights improving disease management. Therefore, useful method routine surveying pathogen-free plants.