A fast and simple assay to quantify bacterial leukotoxin activity

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 standardiz...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Oppermann,Tobias, Schwarz,Stefan, Busse,Nadine, Czermak,Peter
Lenguaje:	English
Publicado:	Pontificia Universidad Católica de Valparaíso 2016
Materias:	ATP assay Bacterial pathogen Bovine respiratory disease Cattle industry Critical parameters Lipopolysaccharides Mannheimia haemolytica Microbial biotechnology Polymyxin B Quantification Virulence
Acceso en línea:	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582016000600006
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:scielo:S0717-34582016000600006
record_format	dspace
spelling	oai:scielo:S0717-345820160006000062017-01-25A fast and simple assay to quantify bacterial leukotoxin activityOppermann,TobiasSchwarz,StefanBusse,NadineCzermak,Peter ATP assay Bacterial pathogen Bovine respiratory disease Cattle industry Critical parameters Lipopolysaccharides Mannheimia haemolytica Microbial biotechnology Polymyxin B Quantification Virulence 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.info:eu-repo/semantics/openAccessPontificia Universidad Católica de ValparaísoElectronic Journal of Biotechnology v.19 n.6 20162016-11-01text/htmlhttp://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582016000600006en10.1016/j.ejbt.2016.10.001
institution	Scielo Chile
collection	Scielo Chile
language	English
topic	ATP assay Bacterial pathogen Bovine respiratory disease Cattle industry Critical parameters Lipopolysaccharides Mannheimia haemolytica Microbial biotechnology Polymyxin B Quantification Virulence
spellingShingle	ATP assay Bacterial pathogen Bovine respiratory disease Cattle industry Critical parameters Lipopolysaccharides Mannheimia haemolytica Microbial biotechnology Polymyxin B Quantification Virulence Oppermann,Tobias Schwarz,Stefan Busse,Nadine Czermak,Peter A fast and simple assay to quantify bacterial leukotoxin activity
description	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.
author	Oppermann,Tobias Schwarz,Stefan Busse,Nadine Czermak,Peter
author_facet	Oppermann,Tobias Schwarz,Stefan Busse,Nadine Czermak,Peter
author_sort	Oppermann,Tobias
title	A fast and simple assay to quantify bacterial leukotoxin activity
title_short	A fast and simple assay to quantify bacterial leukotoxin activity
title_full	A fast and simple assay to quantify bacterial leukotoxin activity
title_fullStr	A fast and simple assay to quantify bacterial leukotoxin activity
title_full_unstemmed	A fast and simple assay to quantify bacterial leukotoxin activity
title_sort	fast and simple assay to quantify bacterial leukotoxin activity
publisher	Pontificia Universidad Católica de Valparaíso
publishDate	2016
url	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582016000600006
work_keys_str_mv	AT oppermanntobias afastandsimpleassaytoquantifybacterialleukotoxinactivity AT schwarzstefan afastandsimpleassaytoquantifybacterialleukotoxinactivity AT bussenadine afastandsimpleassaytoquantifybacterialleukotoxinactivity AT czermakpeter afastandsimpleassaytoquantifybacterialleukotoxinactivity AT oppermanntobias fastandsimpleassaytoquantifybacterialleukotoxinactivity AT schwarzstefan fastandsimpleassaytoquantifybacterialleukotoxinactivity AT bussenadine fastandsimpleassaytoquantifybacterialleukotoxinactivity AT czermakpeter fastandsimpleassaytoquantifybacterialleukotoxinactivity
_version_	1718441937834344448

A fast and simple assay to quantify bacterial leukotoxin activity

Ejemplares similares