This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Supplemental material
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
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 Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by El-Hajj, H. H.
Right arrow Articles by Alland, D.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by El-Hajj, H. H.
Right arrow Articles by Alland, D.

 Previous Article  |  Next Article 

Journal of Clinical Microbiology, April 2009, p. 1190-1198, Vol. 47, No. 4
0095-1137/09/$08.00+0     doi:10.1128/JCM.02043-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Use of Sloppy Molecular Beacon Probes for Identification of Mycobacterial Species {triangledown} ,{dagger}

Hiyam H. El-Hajj,1 Salvatore A. E. Marras,1,2 Sanjay Tyagi,1,3 Elena Shashkina,1 Mini Kamboj,4 Timothy E. Kiehn,5 Michael S. Glickman,4 Fred Russell Kramer,1,2* and David Alland3

Public Health Research Institute, Department of Microbiology and Molecular Genetics,1 Department of Medicine,2 New Jersey Medical School, University of Medicine and Dentistry of New Jersey, Newark, New Jersey, and Division of Infectious Diseases,3 Department of Clinical Laboratories,4 Memorial Sloan-Kettering Cancer Center, New York, New York5

Received 21 October 2008/ Returned for modification 16 December 2008/ Accepted 15 January 2009

We report here the use of novel "sloppy" molecular beacon probes in homogeneous PCR screening assays in which thermal denaturation of the resulting probe-amplicon hybrids provides a characteristic set of amplicon melting temperature (Tm) values that identify which species is present in a sample. Sloppy molecular beacons possess relatively long probe sequences, enabling them to form hybrids with amplicons from many different species despite the presence of mismatched base pairs. By using four sloppy molecular beacons, each possessing a different probe sequence and each labeled with a differently colored fluorophore, four different Tm values can be determined simultaneously. We tested this technique with 27 different species of mycobacteria and found that each species generates a unique, highly reproducible signature that is unaffected by the initial bacterial DNA concentration. Utilizing this general paradigm, screening assays can be designed for the identification of a wide range of species.

* Corresponding author. Mailing address: Public Health Research Institute, 225 Warren Street, Newark, NJ 07103. Phone: (973) 854-3370. Fax: (973) 854-3371. E-mail: Fred.Kramer{at}umdnj.edu

{triangledown} Published ahead of print on 26 January 2009.

{dagger} Supplemental material for this article may be found at http://jcm.asm.org/.

Journal of Clinical Microbiology, April 2009, p. 1190-1198, Vol. 47, No. 4
0095-1137/09/$08.00+0     doi:10.1128/JCM.02043-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Vallee-Belisle, A., Ricci, F., Plaxco, K. W. (2009). Thermodynamic basis for the optimization of binding-induced biomolecular switches and structure-switching biosensors. Proc. Natl. Acad. Sci. USA 106: 13802-13807 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»