Cloning, expression, and purification of the hemolysin/cytolysin (HlyE antigen) from Salmonella enterica serovar Typhi: potential application for immunoassay development

Eugene Boon Beng Ong; Amy Amilda Anthony; Aziah Ismail; Asma Ismail; Theam Soon Lim

doi:10.1016/j.diagmicrobio.2013.05.010

Cloning, expression, and purification of the hemolysin/cytolysin (HlyE antigen) from Salmonella enterica serovar Typhi: potential application for immunoassay development

Diagn Microbiol Infect Dis. 2013 Sep;77(1):87-9. doi: 10.1016/j.diagmicrobio.2013.05.010. Epub 2013 Jun 20.

Authors

Eugene Boon Beng Ong¹, Amy Amilda Anthony, Aziah Ismail, Asma Ismail, Theam Soon Lim

Affiliation

¹ Institute for Research in Molecular Medicine, Universiti Sains Malaysia, 11800 Penang, Malaysia.

PMID: 23790417
DOI: 10.1016/j.diagmicrobio.2013.05.010

Abstract

The hemolysin (HlyE) protein of Salmonella enterica serovar Typhi was reported to be antigenic. This work describes the cloning, expression, and purification of a hexahistidine-tagged HlyE protein under native conditions. Immunoblot analysis and a competitive enzyme-linked immunosorbent assay using sera from typhoid patients showed the presence of HlyE-specific antibodies in circulation.

Keywords: Antigen; ELISA; HlyE; Immunoblot; Salmonella; Typhi.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Antibodies, Bacterial / blood*
Antigens, Bacterial* / genetics
Antigens, Bacterial* / isolation & purification
Clinical Laboratory Techniques / methods*
Cloning, Molecular
Enzyme-Linked Immunosorbent Assay / methods
Gene Expression
Hemolysin Proteins* / genetics
Hemolysin Proteins* / isolation & purification
Humans
Immunoblotting / methods
Recombinant Proteins / genetics
Recombinant Proteins / isolation & purification
Salmonella typhi / genetics
Salmonella typhi / immunology*
Typhoid Fever / diagnosis*

Substances

Antibodies, Bacterial
Antigens, Bacterial
Hemolysin Proteins
HlyE protein, Salmonella enterica
Recombinant Proteins