RESUMO
Crimean-Congo hemorrhagic fever virus (CCHFV) is a geographically widespread RNA virus with a high degree of genomic diversity that complicates sequence-based diagnostics. Here, we sequenced eight CCHFV strains for improved assay design and deposition into FDA-ARGOS, the FDA's pathogen database for development and verification of next generation sequencing assays.
RESUMO
A novel recombinant single-chain fragment variable (scFv) antibody against western equine encephalitis (WEE) virus has been previously constructed and partially characterized. The RS10B5huFc antibody was made by fusing an anti-WEE scFv to a human heavy-chain IgG1 constant region. The RS10B5huFc antibody was functional in binding to WEE virus in enzyme-linked immunosorbent assays (ELISAs), and the Fc domain of the antibody was capable of effector functions, such as binding to protein G and human complement. In this study, the RS10B5huFc antibody was further characterized by BIAcore analyses and was found to possess a binding affinity to a WEE virus epitope (K[D] = 9.14 x 10(-6) M), 4.5-fold lower than its parental mouse monoclonal antibody (MAb) 10B5 E7E2 (K[D] = 2 x 10(-6) M). No cross-reactivity was found between the RS10B5huFc antibody and three other alphaviruses (Sindbis virus [SIN], Venezuelan equine encephalitis [VEE] virus, and eastern equine encephalitis [EEE] virus). Pharmacokinetics studies showed that the RS10B5huFc antibody (free and encapsulated) was found to be retained in the lungs of mice for greater than 48 h when administered intranasally. In contrast, when administered intramuscularly to mice, the RS10B5huFc antibody was not detected in the lungs and only found in the liver and kidneys.
Assuntos
Anticorpos Antivirais/administração & dosagem , Vírus da Encefalite Equina do Oeste/imunologia , Fragmentos de Imunoglobulinas/administração & dosagem , Região Variável de Imunoglobulina/administração & dosagem , Proteínas Recombinantes de Fusão/administração & dosagem , Administração Intranasal , Alphavirus/imunologia , Animais , Anticorpos Antivirais/metabolismo , Anticorpos Antivirais/farmacologia , Especificidade de Anticorpos , Reações Cruzadas , Composição de Medicamentos , Fragmentos de Imunoglobulinas/metabolismo , Fragmentos de Imunoglobulinas/farmacologia , Região Variável de Imunoglobulina/metabolismo , Região Variável de Imunoglobulina/farmacologia , Injeções Intramusculares , Lipossomos , Camundongos , Camundongos Endogâmicos BALB C , Especificidade de Órgãos , Proteínas Recombinantes de Fusão/farmacocinética , Distribuição TecidualRESUMO
OBJECTIVE: To evaluate the effectiveness of a commercially available ELISA kit for detecting antibodies against Borrelia burgdorferi in dogs. SAMPLE POPULATION: Banked sera from 440 military working dogs were used for serologic analyses. PROCEDURE: Serum samples were analyzed for antibodies against B burgdorferi by use of a commercially available ELISA and subsequently by western blot analysis as a confirmatory test. RESULTS: Results from the ELISA indicated that 89 (20%) samples were positive for exposure to B burgdorferi or canine Lyme disease vaccine, and 351 (80%) were negative. Follow-up testing by western blot analysis indicated that results for 109 (25%) samples were positive and 331 (75%) were negative for exposure. All samples that had positive results on the ELISA also had positive results on western blot analysis (true positives). Of the 351 samples that had negative results on the ELISA, only 331 had negative results on western blot analysis (true negatives). The remaining 20 samples had positive results on western blot analysis. By use of a standard 2 x 2 table, it was determined that the ELISA had a sensitivity of 82%, specificity of 100%, positive predictive value of 100%, and negative predictive value of 94%. CONCLUSIONS AND CLINICAL RELEVANCE: The commercial ELISA kit evaluated in this study appeared to lack adequate sensitivity for detecting all potential cases of borreliosis in dogs. The ELISA was also unable to discriminate natural exposure from exposure attributable to vaccination, which could complicate interpretation of positive results and treatment of dogs with clinical signs.
Assuntos
Anticorpos Antibacterianos/sangue , Grupo Borrelia Burgdorferi/imunologia , Doenças do Cão/diagnóstico , Ensaio de Imunoadsorção Enzimática/veterinária , Doença de Lyme/veterinária , Animais , Western Blotting , Cães , Ensaio de Imunoadsorção Enzimática/normas , Estudos de Avaliação como Assunto , Doença de Lyme/diagnóstico , Kit de Reagentes para Diagnóstico/normas , Kit de Reagentes para Diagnóstico/veterinária , Reprodutibilidade dos Testes , Sensibilidade e EspecificidadeAssuntos
Academias e Institutos/história , Patologia Clínica/história , Academias e Institutos/organização & administração , História do Século XX , Hospitais de Ensino/história , Patologia Clínica/organização & administração , Pesquisa/história , Pesquisa/organização & administração , Austrália do SulRESUMO
In spite of rigorous government programs for control of the pricing and dissemination of pharmaceutical products in Australia, the list of new drugs continues to grow and prices to increase. To regain control over drug usage at Royal Adelaide Hospital, the Hospital Drug Committee developed a rating method that judged drugs on the basis of their cost-benefit to patients. The ratio of a total quality score to a total cost score becomes the determinant of additions to the hospital formulary. The background for the Australian approach to pharmaceuticals and the new evaluation technique at the teaching hospital are described in this report.