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


     

Clinical Chemistry 53: 1593-1600, 2007. First published July 27, 2007; 10.1373/clinchem.2007.085472
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow 085472.Supplemental Data
Right arrow All Versions of this Article:
clinchem.2007.085472v1
53/9/1593    most recent
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 Google Scholar
Google Scholar
Right arrow Articles by Winn-Deen, E. S.
Right arrow Articles by Radich, J. P.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Winn-Deen, E. S.
Right arrow Articles by Radich, J. P.
Related Collections
Right arrow Molecular Diagnostics and Genetics
Right arrow Automation and Analytical Techniques
(Clinical Chemistry. 2007;53:1593-1600.)
© 2007 American Association for Clinical Chemistry, Inc.

Molecular Diagnostics and Genetics

Development of an Integrated Assay for Detection of BCR-ABL RNA

Emily S. Winn-Deen1,a, Bret Helton1, Reuel Van Atta1, Wendy Wong1, Jeffrey Peralta1, James Wang1, Gregory J. Tsongalis2, Dorothy Belloni2, David Chan2, James R. Eshleman3, Christopher D. Gocke3, Zsolt Jobbagy3, Lan Beppu4 and Jerald P. Radich4

1 Cepheid, Sunnyvale, CA.
2 Dartmouth-Hitchcock Medical Center, Lebanon, NH.
3 Molecular Diagnostics Laboratory, Johns Hopkins Medical Institutions, Baltimore, MD.
4 Fred Hutchinson Cancer Research Center, Seattle, WA.

aAddress correspondence to this author at: Cepheid, 904 Caribbean Dr., Sunnyvale, CA 94089-1189. Fax 408-541-6439; e-mail Emily.Winn-Deen{at}cepheid.com.

Background: Current practice guidelines for managing patients with chronic myelogenous leukemia (CML) call for monitoring BCR-ABL transcript concentrations with a quantitative reverse transcription–PCR (qRT-PCR) assay. Because the available laboratory-developed assays lack consensus on the appropriate design, reporting of results, and reference intervals, we developed and evaluated an integrated BCR-ABL assay that yields standardized results for any laboratory and can be performed by technicians with no specialized training.

Methods: We used the Cepheid Xpert® BCR-ABL Monitor assay to measure both BCR-ABL and ABL (endogenous control) transcripts in blood samples from CML patients and healthy individuals. The assay involves 8 manual pipetting steps, fully automated nucleic acid purification, a nested qRT-PCR step, and data analysis.

Results: The BCR-ABL assay requires approximately 2 h 20 min and covers a 5-log concentration range with a lower detection limit for the BCR-ABL:ABL ratio of approximately 0.005%. Assay results were negative for 100% of the 56 known CML-negative samples (12 patients with other hematologic disorders and 44 healthy blood donors). Testing of CML-positive patients undergoing disease monitoring showed 85% agreement with negative results (17 of 20) and 100% agreement with positive results (26 of 26). An imprecision/portability study revealed no differences in performance between sites, days, instruments, and operators.

Conclusions: The Xpert BCR-ABL Monitor assay provides a robust and reproducible alternative to laboratory-developed assays. Its ease of use may allow more laboratories to offer BCR-ABL testing for patients, and the short assay time enables same-day results for treating physicians.






HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
Copyright © 2007 by the American Association for Clinical Chemistry.