Document Detail

A novel homogeneous immunoassay for anthrax detection based on the AlphaLISA method: detection of B. anthracis spores and protective antigen (PA) in complex samples.
MedLine Citation:
PMID:  23388689     Owner:  NLM     Status:  Publisher    
Amplified Luminescent Proximity Homogeneous Assay (AlphaLISA) technology is an energy-transfer-based assay, utilizing singlet oxygen as an energy donor to a fluorescent acceptor. The long singlet oxygen migration distance allows the energy transfer mechanism to go up to ∼200 nm, facilitating flexible and sensitive homogeneous immunoassays. While soluble protein detection using AlphaLISA was previously described, the detection of particles such as bacteria and viruses was not reported. In this work, we show for the first time the implementation of the AlphaLISA technology for the detection of a particulate antigen, i.e., Bacillus anthracis spores. Here, we show that an efficient particle immunoassay requires a high acceptor-to-donor ratio (>4:1). The results suggested that the high acceptor/donor ratio is required to avoid donor aggregation ("islands") on the spore surface, hence facilitating donor/acceptor interaction. The developed assay enabled the detection of 10(6) spores/mL spiked in PBS. We also demonstrate the development of a highly sensitive AlphaLISA assay for the detection of the main toxin component of anthrax, protective antigen (PA). The assay enabled the detection of 10 and 100 pg/mL PA in buffer and spiked naïve rabbit sera, respectively, and was successfully implemented in sera of anthrax-infected rabbits. To summarize, this study demonstrates that AlphaLISA enables detection of anthrax spores and toxin, utilizing short homogeneous assays. Moreover, it is shown for the first time that this technology facilitates the detection of particulate entities and might be suitable for the detection of other bacteria or viruses.
Adva Mechaly; Noam Cohen; Shay Weiss; Eran Zahavy
Related Documents :
22750039 - Novel taqman® pcr for detection of leptospira species in urine and blood: pit-falls of...
22890869 - High-performance liquid chromatography method for determination of hydrogen peroxide in...
22281679 - Sequential injection ionic liquid dispersive liquid-liquid microextraction for thallium...
22364669 - Optimization of an improved single-column chromatographic process for the separation of...
22702459 - Development of a reverse transcription loop-mediated isothermal amplification assay for...
22349469 - Influence of the ionic liquid/gas surface on ionic liquid chemistry.
10869689 - High-resolution gas-chromatographic analysis of the secondary metabolites obtained by s...
3856229 - Differences between two fractions of glycosaminoglycans of the corneal stroma in their ...
18258499 - Quantification of cationic anti-malaria agent methylene blue in different human biologi...
Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2013-2-7
Journal Detail:
Title:  Analytical and bioanalytical chemistry     Volume:  -     ISSN:  1618-2650     ISO Abbreviation:  Anal Bioanal Chem     Publication Date:  2013 Feb 
Date Detail:
Created Date:  2013-2-7     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  101134327     Medline TA:  Anal Bioanal Chem     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
Department of Infectious Diseases, Israel Institute for Biological Research, Ness Ziona, 74100, Israel.
Export Citation:
APA/MLA Format     Download EndNote     Download BibTex
MeSH Terms

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

Previous Document:  Simultaneous serum desalting and total protein determination by macroporous reversed-phase chromatog...
Next Document:  Use of electrophoretic techniques and MALDI-TOF MS for rapid and reliable characterization of bacter...