Document Detail


Increasing the sensitivity of Listeria monocytogenes assays: evaluation using ELISA and amperometric detection.
MedLine Citation:
PMID:  10605890     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
An immunosensor for the detection of Listeria monocytogenes was developed. ELISA and amperometric studies were run in parallel to develop a more sensitive and rapid assay for the bacterium. Conditions for the immunosensor were primarily characterised using ELISA. A direct sandwich assay was employed and the affinities of two polyclonal (goat and rabbit) and one monoclonal (mouse) anti-L. monocytogenes antibodies were compared using this format. Owing to low sensitivity being obtained with all antibodies, biotin-avidin amplification and an indirect sandwich assay were employed. The system was then transferred to screen-printed electrodes (SPEs), the primary antibody being immobilised by cross-linking with 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide, and the mode of detection being amperometric. Various parameters (limit of detection, working range, incubation time, cross-reactivity) of the systems were characterised. The effect of direct incubation in milk is also discussed. The final immunosensor had a working range of 1 x 10(6)-1 x 10(3) cells ml-1 and a detection limit of 9 x 10(2) cells ml-1. The assay took about 3.5 h to complete.
Authors:
E L Crowley; C K O'Sullivan; G G Guilbault
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  The Analyst     Volume:  124     ISSN:  0003-2654     ISO Abbreviation:  Analyst     Publication Date:  1999 Mar 
Date Detail:
Created Date:  2000-01-11     Completed Date:  2000-01-11     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  0372652     Medline TA:  Analyst     Country:  ENGLAND    
Other Details:
Languages:  eng     Pagination:  295-9     Citation Subset:  IM    
Affiliation:
Laboratory of Sensor Development, National University of Ireland Cork, Ireland.
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MeSH Terms
Descriptor/Qualifier:
Biosensing Techniques
Enzyme-Linked Immunosorbent Assay
Humans
Listeria monocytogenes / isolation & purification*

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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