Clinical Chemistry Link to Randox Laboratories Web Site
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
QUICK SEARCH:   [advanced]


     

Clinical Chemistry 49: 407-414, 2003; 10.1373/49.3.407
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Submit an electronic Letter to
the Editor about this paper
Right arrow Alert me when this article is cited
Right arrow Alert me when eLetters are posted
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in ISI Web of Science
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via ISI Web of Science (11)
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Moser, M. J.
Right arrow Articles by Prudent, J. R.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Moser, M. J.
Right arrow Articles by Prudent, J. R.
Related Collections
Right arrow Molecular Diagnostics and Genetics
(Clinical Chemistry. 2003;49:407-414.)
© 2003 American Association for Clinical Chemistry, Inc.

Exploiting the Enzymatic Recognition of an Unnatural Base Pair to Develop a Universal Genetic Analysis System

Michael J. Moser1, David J. Marshall1, Jennifer K. Grenier1, Collin D. Kieffer1, Anthony A. Killeen2, Jerod L. Ptacin1, Craig S. Richmond1, Eric B. Roesch1, Christopher W. Scherrer1, Christopher B. Sherrill1, Cris V. Van Hout1, Sara J. Zanton1 and James R. Prudent1a

1 Eragen Biosciences, Inc., 918 Deming Way, Madison, WI 53717-1944.

2 Department of Pathology, University of Michigan, Ann Arbor, MI 48109.

aAuthor for correspondence. E-mail jprudent{at}eragen.com.

Background: With the invention of the DNA chip, genome-wide analysis is now a reality. Unfortunately, solid-phase detection systems such as the DNA chip suffer from a narrow range in quantification and sensitivity. Today the best methodology for sensitive, wide dynamic range quantification and genotyping of nucleic acids is real-time PCR. However, multiplexed real-time PCR technologies require complicated and costly design and manufacturing of separate detection probes for each new target.

Methods: We developed a novel real-time PCR technology that uses universal energy transfer probes constructed from An Expanded Genetic Information System (AEGIS) for both quantification and genotyping analyses.

Results: RNA quantification by reverse transcription-PCR was linear over four orders of magnitude for the simultaneous analysis of ß-actin messenger RNA and 18S ribosomal RNA. A single trial validation study of 176 previously genotyped clinical specimens was performed by endpoint analysis for factor V Leiden and prothrombin 20210A mutation detection. There was concordance for 173 samples between the genotyping results from Invader® tests and the AEGIS universal energy transfer probe system for both factor V Leiden and prothrombin G20210A. Two prothrombin and one factor V sample gave indeterminate results (no calls).

Conclusion: The AEGIS universal probe system allows for rapid development of PCR assays for nucleic acid quantification and genotyping.




The following articles in journals at HighWire Press have cited this article:


Home page
 Clin. Chem.Home page
D. R. Christensen, L. J. Hartman, B. M. Loveless, M. S. Frye, M. A. Shipley, D. L. Bridge, M. J. Richards, R. S. Kaplan, J. Garrison, C. D. Baldwin, et al.
Detection of Biological Threat Agents by Real-Time PCR: Comparison of Assay Performance on the R.A.P.I.D., the LightCycler, and the Smart Cycler Platforms
Clin. Chem., January 1, 2006; 52(1): 141 - 145.
[Abstract] [Full Text] [PDF]

Home page
 Clin. Chem.Home page
S. C. Johnson, D. J. Marshall, G. Harms, C. M. Miller, C. B. Sherrill, E. L. Beaty, S. A. Lederer, E. B. Roesch, G. Madsen, G. L. Hoffman, et al.
Multiplexed Genetic Analysis Using an Expanded Genetic Alphabet
Clin. Chem., November 1, 2004; 50(11): 2019 - 2027.
[Abstract] [Full Text] [PDF]

Home page
 Clin. Chem.Home page
A. M. Rickert, H. Lehrach, and S. Sperling
Multiplexed Real-Time PCR Using Universal Reporters
Clin. Chem., September 1, 2004; 50(9): 1680 - 1683.
[Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
S. C. Johnson, C. B. Sherrill, D. J. Marshall, M. J. Moser, and J. R. Prudent
A third base pair for the polymerase chain reaction: inserting isoC and isoG
Nucleic Acids Res., March 29, 2004; 32(6): 1937 - 1941.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
M. J. Moser and J. R. Prudent
Enzymatic repair of an expanded genetic information system
Nucleic Acids Res., September 1, 2003; 31(17): 5048 - 5053.
[Abstract] [Full Text] [PDF]


HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
Copyright © 2003 by the American Association for Clinical Chemistry.