Clinical Chemistry AACC Online Job Center
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
QUICK SEARCH:   [advanced]


     

Clinical Chemistry 52: 430-437, 2006. First published January 12, 2006; 10.1373/clinchem.2005.061259
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow All Versions of this Article:
clinchem.2005.061259v1
52/3/430    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 ISI Web of Science (1)
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Zehentner, B. K.
Right arrow Articles by Loken, M. R.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Zehentner, B. K.
Right arrow Articles by Loken, M. R.
Related Collections
Right arrow Molecular Diagnostics and Genetics
Right arrow Cancer Diagnostics (since 2002)
(Clinical Chemistry. 2006;52:430-437.)
© 2006 American Association for Clinical Chemistry, Inc.

Cancer Diagnostics

Minimal Disease Detection and Confirmation in Hematologic Malignancies: Combining Cell Sorting with Clonality Profiling

Barbara K. Zehentner1,a, Wayne Fritschle1, Tess Stelzer1, Keely M. Ghirardelli1, Kimberly Hunter1, Collette Wentzel1, Richard Bennington1, Christian L. Hansen3, David Myerson2, Michael Kalnoski1, Denise A. Wells1 and Michael R. Loken1

1 HematoLogics, Inc., Seattle, WA.
2 Fred Hutchinson Cancer Research Center, Seattle, WA.
3 Community Hospital of the Monterey Peninsula, Pathology, Monterey, CA.

aAddress correspondence to this author at: HematoLogics, Inc., Molecular Analysis, 113 1st Ave. North, Seattle, WA 98109. Fax 206-223-5550; e-mail Barbara{at}hematologics.com.

Background: In this study we demonstrate the technical application of flow cytometry and cell sorting combined with gene-rearrangement clonality profiling to detect and confirm minimal disease in 2 leukemia and 2 lymphoma cases.

Methods: Specimens with low percentages (0.05%–5%) of abnormal lymphoid populations were identified by flow cytometry. The abnormal lymphoid populations were sorted by flow cytometry, and the purified tumor populations along with unsorted fractions were subsequently analyzed for the presence of clonal gene rearrangements by PCR and fluorescence-based capillary electrophoresis fragment analysis.

Results: In 3 cases, distinct clonality profiles could be detected in the purified tumor cell fraction, and suspicious amplicons of identical sizes were detected among the polyclonal backgrounds in the unsorted specimens. For 1 patient, a monoclonal signal was detected in the sorted tumor cell fraction but not in the unseparated bone marrow specimen containing 0.05% abnormal lymphoblasts. A subsequent bone marrow specimen containing 4.8% recurring leukemia cells tested positive with a clonality profile that matched the previous profile in the sorted cell population.

Conclusions: The described method integrating 2 technologies allows genotypic confirmation of an aberrant population detected by immunophenotype to increase diagnostic certainty. This strategy provides a sensitive tool for disease monitoring without the need for patient-specific primer design and assay optimization required for quantitative PCR analysis.






HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
Copyright © 2006 by the American Association for Clinical Chemistry.