72 research outputs found
APOSTEL 2.0 recommendations for reporting quantitative optical coherence tomography studies
OBJECTIVE: To update the consensus recommendations for reporting of quantitative optical coherence tomography (OCT) study results, thus revising the previously published Advised Protocol for OCT Study Terminology and Elements (APOSTEL) recommendations. METHODS: To identify studies reporting quantitative OCT results, we performed a PubMed search for the terms âquantitativeâ and âoptical coherence tomographyâ from 2015 to 2017. Corresponding authors of the identified publications were invited to provide feedback on the initial APOSTEL recommendations via online surveys following the principle of a modified Delphi method. The results were evaluated and discussed by a panel of experts, and changes to the initial recommendations were proposed. A final survey was recirculated among the corresponding authors to obtain a majority vote on the proposed changes. RESULTS: One hundred sixteen authors participated in the surveys, resulting in 15 suggestions, of which 12 were finally accepted and incorporated into an updated 9-point-checklist. We harmonized the nomenclature of the outer retinal layers, added the exact area of measurement to the description of volume scans; we suggested reporting device-specific features. We advised to address potential bias in manual segmentation or manual correction of segmentation errors. References to specific reporting guidelines and room light conditions were removed. The participantsâ consensus with the recommendations increased from 80% for the previous APOSTEL version to greater than 90%. CONCLUSIONS: The modified Delphi method resulted in an expert-led guideline (evidence class III, GRADE criteria) concerning study protocol, acquisition device, acquisition settings, scanning protocol, fundoscopic imaging, post-acquisition data selection, post-acquisition analysis, nomenclature and abbreviations, and statistical approach. It will still be essential to update these recommendations to new research and practices regularly
Primers based on the rpf gene region provide improved detection of Xanthomonas axonopodis pv. citri in naturally and artificially infected citrus plants
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Single chain antibodies against âCa. Liberibacter asiaticusâ
Antibodies are widely used as microbiological reagents, but antibodies that recognize âCa. Liberibacter asiaticusâ are generally lacking. We have developed and applied immunization and affinity screening methods to create a primary library of recombinant single chain variable fragment (scFv) antibodies in an M13 vector, pKM19. The antibody population is enriched for antibodies that bind antigens of âCa. Liberibacter asiaticusâ and Diaphorina citri. The primary library has more than 107 unique antibodies and the genes that encode them. We have screened this library of antibodies for antibodies that bind to specifically chosen proteins that are present on the surface of âCa. Liberibacter asiaticusâ. These proteins were used as âbaitâ for affinity-based selection of scFvs that bind to the major outer membrane protein, OmpA; the polysaccharide capsule expressing protein KpsF; a protein component of the type IV pilus (CapF); and two flagellar proteins FlhA and FlgI. These scFvs have been used in ELISA and dot blot assays against purified protein antigens and âCa. Liberibacter asiaticusâ infected plant extracts. We also have isolated scFv that bind to surface exposed portions of the TolC proteins and of a protein called InvA. These proteins may have critical roles in pathogenicity. Thus far, screening of these scFvs is more efficient when using phage bound, rather than soluble scFvs. We have demonstrated a technology to produce antibodies and select at will and against any protein target encoded by âCa. Liberibacter asiaticusâ. Future applications will include advanced diagnostic methods for huanglongbing and the development of immune labeling reagents for in planta applications
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