By Hayward's classification method, based on carbon source utilisation, 19 of the 62 R. solanacearum based on their cultural characteristics on TTC medium, tomato pathogenicity bioassay, carbon source utilisation patterns and a specific PCR-based assay. Of these, 62 strains were identified as R. potato, tomato and pepper plants and potato tubers) at various sites in Ethiopia.
#BACTERIAL BLIGHT OF GERNAIUM PORTABLE#
These two independent, portable systems have the potential to facilitate Rs r3b2 detection at the ports of entry and in field settings.Įighty one isolates of Ralstonia solanacearum -like bacteria on triphenyl tetrazolium chloride (TTC) medium were collected from different Solanaceae crops (i.e.
Moreover, the Rs MAb-coupled sensors are sensitive to Rs r3b2 after three months of room temperature storage, although the detection sensitivity was reduced by 28%. The Rs MAb/ BLItz ® system has comparable rapidity and sensitivity to the commercial ImmunoStrip ® with the advantage of higher r3b2 specificity. Additionally, we demonstrate that the BLI-based BLItz ® instrument with the Forsite Rs MAb, which is more reactive to UW551 strain than other Rs species complex members, can detect as low as 2.5 × 10 5 CFU/ml of UW551 in spiked geranium extract in a few minutes. The RsSA2/POCKIT™ system can specifically detect heat-inactivated Rs r3b2 strains, and such cells in spiked geranium stem sections, as well as in UW551-infected symptomatic and asymptomatic geranium plants. Our data indicate that combining the palm sized POCKIT™ with the previously published TaqMan-based qPCR primers and probe set RsSA2, which targets a non-phage r3b2-unique sequence annotated as a probable n-6 adenine-specific DNA methylase, can detect as low as 10 CFU of the r3b2 strain UW551 in 32 minutes. We report here the development of two portable platforms for r3b2 detection, the TaqMan probe qPCR (quantitative, real-time polymerase chain reaction)-based POCKIT™ and the Rs monoclonal antibody (MAb)-coupled sensor BLI (bio-layer interferometry)-based BLItz ®. Rapid, sensitive and accurate detection methods are urgently needed. Ralstonia solanacearum (Rs) race 3 biovar 2 (r3b2) is designated as a quarantine pathogen in many countries and additionally as a Select Agent in the United States.
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