| Autor: |
Lee, J.J., Hsieh, C.L., Widman, J., Mingala, C., Ardeza Villanueva, M., Feng, H., Divers, T., Chang, Y.‐F. |
| Zdroj: |
Equine Veterinary Journal; Sep2019, Vol. 51 Issue 5, p669-673, 5p |
| Abstrakt: |
Summary: Background: Current serological tests cannot discriminate between bactericidal Borrelia burgdorferi antibodies from others that are merely a response to Borrelia antigenic stimulation. Objective: To develop a sensitive and convenient luminescence‐based serum bactericidal assay (L‐SBA) to identify serum borreliacidal activity. Study design: Prospective validation study and method comparison. Methods: Serum samples were obtained either from archives of the Animal Health Diagnostic Center at Cornell University (N = 7) or from a vaccination trial (N = 238). Endogenous complement‐inactivated serum sample was incubated with exogenic complement and B. burgdorferi ML23 pBBE22luc, which is able to process luciferin with luciferase and produce luminescence in viable Borrelia. After incubation, a light signal can be detected by using a luminometer to calculate the borreliacidal antibody titre. Results: Components of the reaction mixture including spirochetes and complement from various sources and concentrations were tested to identify a reliable recipe for our complement‐mediated L‐SBA. We also applied this L‐SBA on measuring bactericidal antibody activities and calculated the half inhibitory concentration (IC50) of serum samples from clinical collections. Furthermore, we analysed the L‐SBA titres and anti‐outer surface protein A (OspA) antibody levels from vaccinated horses using the multiplex assays and found that there is a relationship between results generated using these two different assays. The increases of L‐SBA titres correlated with increases of anti‐OspA antibody titre in sera (r = 0.423). Main limitations: Immunoreactivity of commercial complement may differ from different batches. Clinical protection of borreliacidal antibody levels has not been determined. Conclusions: The L‐SBA provided a sensitive and easy‐operating platform for the evaluation of bactericidal antibody to B. burgdorferi, and we anticipated L‐SBA would function well as an evaluation tool of vaccine efficiency in the future. [ABSTRACT FROM AUTHOR] |
| Databáze: |
Complementary Index |
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