Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: March 15, 2010
Publication Date: June 1, 2010
Citation: Yuan, Q., Jordan, R.L., Bohannon, R., Brlansky, R.H., Minenkova, O., Hartung, J.S. 2010. Development of conventional monoclonal antibody and recombinant antibody (scFv) against Candidatus Liberibacter asiaticus. Phytopathology 100(6):S43.
A non-culturable member of the alpha-proteobacteria, ‘Ca. Liberibacter asiaticus’, is consistently associated with huanglongbing (HLB) disease. This bacterium is transmitted by citrus psyllids and grows systemically in infected citrus phloem tissues. Control of the disease requires effective, convenient and inexpensive methods for detection of the pathogen in infected plants and insects. Antibodies are the most widely used tool to detect pathogens, and they are also uniquely useful as experimental reagents. Therefore antibodies against the HLB pathogens would greatly aide detection of the pathogen and eventual control of this disease. Extracts of psyllids fed on HLB infected citrus in Florida were assayed individually for ‘Ca. Liberibacter asiaticus’ by q-PCR. Extracts with more than 108 ‘Ca. Liberibacter asiaticus’/100ul were used to immunize BALB/C mice. Monoclonal antibodies were made by spleen/myeloma fusion and screened against both plant extracts containing ‘Ca. Liberibacter asiaticus’ and against outer membrane protein (OMP) purified from Escherichia coli cells expressing the cloned gene. A recombinant library of scFv antibodies also was made using the phage vector pKM19 to clone cDNAs prepared from the mRNA isolated from the mouse spleens. This scFv library in recombinant phage is currently being screened against extracts of plants infected with high concentrations of ‘Ca. Liberibacter asiaticus’ as well as against purified outer membrane protein from ‘Ca. Liberibacter asiaticus’.