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This Article Full Text Full Text (PDF) 085472.Supplemental Data All Versions of this Article: clinchem.2007.085472v1 53/9/1593    most recent Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in Web of Science Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Web of Science (2) Citing Articles via Google Scholar Google Scholar Articles by Winn-Deen, E. S. Articles by Radich, J. P. Search for Related Content PubMed PubMed Citation Articles by Winn-Deen, E. S. Articles by Radich, J. P. Related Collections Molecular Diagnostics and Genetics Automation and Analytical Techniques

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

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.

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				S. Branford, L. Fletcher, N. C. P. Cross, M. C. Muller, A. Hochhaus, D.-W. Kim, J. P. Radich, G. Saglio, F. Pane, S. Kamel-Reid, et al. Desirable performance characteristics for BCR-ABL measurement on an international reporting scale to allow consistent interpretation of individual patient response and comparison of response rates between clinical trials Blood, October 15, 2008; 112(8): 3330 - 3338. [Abstract] [Full Text] [PDF]