Skip Navigation

Nucleic Acids Research 2005 33(2):e13; doi:10.1093/nar/gni015
This Article
Right arrow Full Text Freely available
Right arrow Print PDF (2781K) Freely available
Right arrow Screen PDF (505K) Freely available
Right arrow Supplementary Material
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrow Commercial Re-use Guidelines
for Open Access NAR Content
Google Scholar
Right arrow Articles by Horejsh, D.
Right arrow Articles by Capobianchi, M. R.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Horejsh, D.
Right arrow Articles by Capobianchi, M. R.
Related Collections
Right arrow DNA characterisation
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

Published online 19 January 2005

© 2005, the authors Nucleic Acids Research, Vol. 33 No. 2 © Oxford University Press 2005; all rights reserved
The online version of this article has been published under an open access model. Users are entitled to use, reproduce, disseminate, or display the open access version of this article for non-commercial purposes provided that: the original authorship is properly and fully attributed; the Journal and Oxford University Press are attributed as the original place of publication with the correct citation details given; if an article is subsequently reproduced or disseminated not in its entirety but only in part or as a derivative work this must be clearly indicated. For commercial re-use permissions, please contact journals.permissions{at}oupjournals.org.

Methods Online

A molecular beacon, bead-based assay for the detection of nucleic acids by flow cytometry

Douglas Horejsh*, Federico Martini, Fabrizio Poccia, Giuseppe Ippolito, Antonino Di Caro and Maria R. Capobianchi

National Institute for Infectious Diseases ‘Lazzaro Spallanzani’–IRCCS Rome, Italy

*To whom correspondence should be addressed at INMI L. Spallanzani, Virologia-Baglivi/Del Vecchio, via Portuense 292, 00149 Rome, Italy. Tel: +39 06 551 70907; Fax: +39 06 551 70918; Email: horejsh{at}inmi.it

Received September 2, 2004. Revised October 18, 2004. Accepted December 22, 2004.

Molecular beacons are dual-labelled probes that are typically used in real-time PCR assays, but have also been conjugated with solid matrices for use in microarrays or biosensors. We have developed a fluid array system using microsphere-conjugated molecular beacons and the flow cytometer for the specific, multiplexed detection of unlabelled nucleic acids in solution. For this array system, molecular beacons were conjugated with microspheres using a biotin-streptavidin linkage. A bridged conjugation method using streptavidin increased the signal-to-noise ratio, allowing for further discrimination of target quantitation. Using beads of different sizes and molecular beacons in two fluorophore colours, synthetic nucleic acid control sequences were specifically detected for three respiratory pathogens, including the SARS coronavirus in proof-of-concept experiments. Considering that routine flow cytometers are able to detect up to four fluorescent channels, this novel assay may allow for the specific multiplex detection of a nucleic acid panel in a single tube.


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?


This article has been cited by other articles:


Home page
 J. Clin. Microbiol.Home page
D. J. Marshall, E. Reisdorf, G. Harms, E. Beaty, M. J. Moser, W.-M. Lee, J. E. Gern, F. S. Nolte, P. Shult, and J. R. Prudent
Evaluation of a Multiplexed PCR Assay for Detection of Respiratory Viral Pathogens in a Public Health Laboratory Setting
J. Clin. Microbiol., December 1, 2007; 45(12): 3875 - 3882.
[Abstract] [Full Text] [PDF]


Disclaimer:
Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.