Type

Journal Article

Authors

Richard O'Kennedy
John Quinn
Gary Wyatt
Stephen Hearty
Paul Leonard

Subjects

Microbiology

Topics
molecular weight isolation purification immunology electrophoresis polyacrylamide gel listeria monocytogenes antibodies surface plasmon resonance animals food contamination reproducibility of results enzyme linked immunosorbent assay sensitivity and specificity food microbiology dna bacterial

Development of a surface plasmon resonance-based immunoassay for Listeria monocytogenes. (2005)

Abstract A polyclonal antibody was produced against Internalin B (InlB)-enriched extract and used to develop an inhibition assay to detect Listeria monocytogenes cells in solution using surface plasmon resonance. The gene sequence encoding for the InlB protein was cloned into a Qiagen pQE-60 vector, expressed in Escherichia coli, and purified by immobilized metal affinity chromatography. Protein G-purified anti-InlB-enriched extract polyclonal antibody was incubated with various concentrations of L. monocytogenes cells and subsequently injected over a purified-recombinant InlB (rInlB)-immobilized CM5 sensor chip surface. A decrease in antibody binding response was observed with increasing L. monocytogenes cell concentrations. Intraday and interday assay variability studies were carried out to evaluate precision and reproducibility. The assay had a limit of detection of less than 2 x 10(5) cells per ml and could be successfully reproduced with coefficients of variation of between 2.5 and 7.7%.
Collections Ireland -> Dublin City University -> PubMed

Full list of authors on original publication

Richard O'Kennedy, John Quinn, Gary Wyatt, Stephen Hearty, Paul Leonard

Experts in our system

1
R O'Kennedy
Dublin City University
Total Publications: 197
 
2
Stephen Hearty
Dublin City University
Total Publications: 27
 
3
Paul Leonard
Dublin City University
Total Publications: 27