Data/hora: 05/06/2024 02:46:25

Provedor de dados: | 69
País: | Chile
Título: | A fast and simple assay to quantify bacterial leukotoxin activity
Autores: | Oppermann,Tobias; Schwarz,Stefan; Busse,Nadine; Czermak,Peter.
Data: | 2016-11-01
Ano: | 2016
Palavras-chave: | ATP assay; Bacterial pathogen; Bovine respiratory disease; Cattle industry; Critical parameters; Lipopolysaccharides; Mannheimia haemolytica; Microbial biotechnology; Polymyxin B; Quantification; Virulence.
Resumo: | Background: Mannheimia haemolytica is the primary bacterial pathogen in causing bovine respiratory disease with tremendous annual losses in the cattle industry. The leukotoxin from M. haemolytica is the predominant virulence factor. Several leukotoxin activity assays are available but not standardized regarding sample preparation and cell line. Furthermore, these assays suffer from a high standard error, a prolonged time consumption and often complex sample pretreatments, which is important from the bioprocess engineering point of view. Results: Within this study, an activity assay based on the continuous cell line BL3.1 combined with a commercial available adenosine triphosphate viability assay kit was established. The leukotoxin activity was found to be strongly dependent on the sample preparation. Furthermore, the interfering effect of lipopolysaccharides in the sample could be successfully suppressed by adding polymyxin B. We reached a maximum relative P95 value of 14%, which is more than seven times lower compared to current available assays as well as a time reduction up to 88%. Conclusion: Ultimately, the established leukotoxin activity assay is simple, fast and has a high reproducibility. Critical parameters regarding the sample preparation were characterized and optimized making complex sample purification superfluous.
Tipo: | Journal article
Idioma: | Inglês
Identificador: | http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582016000600006
Editor: | Pontificia Universidad Católica de Valparaíso
Formato: | text/html
Fonte: | Electronic Journal of Biotechnology  v.19 n.6 2016