Validation of colorimetric assay to detect complement-mediated antibody-dependent bactericidal activity against serogroups B and C Neisseria meningitidis |
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Authors: | Rodríguez Tamara Lastre Miriam Cedré Bárbara Fajardo Esther María del Campo Judith Delgado Ileana Sierra Gustavo Pérez Oliver |
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Affiliation: | Department of Immunology, Direction of Bacterial Vaccines and Direction of Control of Quality, Finlay Institute, 27 Ave, b/198 and 202, PO Box 16017, La Lisa, Havana, Cuba. trodriguez@finlay.edu.cu |
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Abstract: | Colorimetric serum bactericidal assay (cSBA), based on the addition of glucose and a pH indicator to the culture medium after the bactericidal reaction, was validated. The precision measured as repeatability, intermediated precision, and reproducibility was determined as a percentage in titer coincidence between replicas >/=50. Moreover the use of the freeze-dried complement was evaluated in comparison to the traditionally stored by freezing. The results were the following: precision: titer +/-1 two-fold dilution (except for the highly positive serum against serogroup B, where there was titer +/-2 dilutions) percentage in titer coincidence: >/=50. The known titer or +/-1 two-fold dilution was found in the sera titrated with the complement either frozen or freeze-dried. Concluding, cSBA showed to be highly precise, allowing also the use of freeze-dried complement which is another important advantage for this kind of assay. |
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