Desarrollo de una prueba inmunocromatográfica para la detección rápida de Bartonella bacilliformis
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AdvisorsDel Valle Mendoza, Juana Mercedes
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Citation1. Aleman M, Martín FD, Espejo W, Claudia A, Mendoza V. Desarrollo de una prueba inmunocromatográfica para la detección rápida de Bartonella bacilliformis [Internet]. Universidad Peruana de Ciencias Aplicadas (UPC); 2017. Available from: http://hdl.handle.net/10757/621857
1. Aleman M, Martín FD, Espejo W, Claudia A, Mendoza V. Desarrollo de una prueba inmunocromatográfica para la detección rápida de Bartonella bacilliformis [Internet]. Universidad Peruana de Ciencias Aplicadas (UPC); 2017. Available from: http://hdl.handle.net/10757/621857
AbstractBackground: Carrión's disease is an important disease that requires a timely diagnosis and management to reduce its morbimortality. Objective: Develop a lateral flow assay for the rapid detection of Bartonella bacilliformis using colloidal gold-labeled rabbit polyclonal antibodies. Materials and methods: Polyclonal antibodies against Bartonella bacilliformis were produced by the immunization of rabbits with GroEL protein purified from Bartonella bacilliformis. Colloidal gold of 40 nm was used for the conjugation process with the rabbit polyclonal antibodies. The analytical sensitivity was evaluated by testing solutions of B. bacilliformis with different concentrations ranging between 1 x 101 to 1 x 105 CFU/mL. Analytical specificity was determined by testing cross-reactivity with microorganisms from different species, including Staphylococcus aureus, Streptococcus pneumoniae, Streptococcus viridans, Chlamydophila pneumoniae, Escherichia coli and Candida spp. Results: The optimal concentrations for the capture antibody and the coating antibody (T-line) were 2 mg / mL and 0.4 mg / mL, respectively. The analytical sensitivity was determined to be between 1x102 CFU/mL and 1x101 CFU/mL. There were no cross-reactions observed with the groups of bacteria used in this study. We determined that the flowing time and volume required for an optimum signal to be generated was 20 minutes and 50 µL, respectively. Conclusions: We developed the first gold-based lateral flow assay for the rapid, sensitive and specific detection of Bartonella bacilliformis using polyclonal antibodies against the protein GroEL, which needs to be validated in future studies.
DescriptionEsta tesis se encuentra en proceso de registro como patente.