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Updated: Aug 8, 2026
A Semi-automated Approach to Preparing Antibody Cocktails for Immunophenotypic Analysis of Human Peripheral Blood
Published on: February 8, 2016
Single step immunophenotyping of acute leukemias not classifiable by standard morphology and cytochemistry: a
R Raimondi1, G Pellizzari, F Rodeghiero
1Department of Hematology, San Bortolo Hospital, Vicenza, Italy.
Insights
Immunophenotyping using a panel of nine monoclonal antibodies aids in diagnosing acute leukemia, especially for cases unclassifiable by standard methods. This approach provides rapid, cost-effective typing for timely therapeutic decisions.
Area of Science:
- Hematology
- Immunology
- Oncology
Background:
- Immunophenotyping is crucial for diagnosing acute leukemia, particularly when morphology and cytochemistry are inconclusive.
- Various immunomarker panels have been proposed to aid in acute leukemia classification.
- Accurate diagnosis is essential for effective treatment strategies.
Purpose of the Study:
- To evaluate the utility of a specific panel of monoclonal antibodies for immunophenotyping acute leukemia.
- To determine if a reduced antibody panel can reliably classify acute leukemias.
- To assess the cost-effectiveness and speed of immunophenotyping for clinical decision-making.
Main Methods:
- Peripheral blood and/or bone marrow samples from 125 patients were analyzed.
- Flow cytometry with single and double fluorescent labeling was employed.
- Reactivity of 17 monoclonal antibodies was initially assessed.
Main Results:
- Out of 125 patients, 75 were classified as acute lymphoblastic leukemia (ALL) (58 B-lineage, 17 T-lineage).
- 33 patients were classified as acute myeloid leukemia (AML), including 2 M7.
- 6.4% were biphenotypic and 8.8% were undifferentiated.
Conclusions:
- A panel of nine monoclonal antibodies (CD2, CD5, CD7, CD10, CD19, CD20, CD13, CD33, CD41) is sufficient for reliable acute leukemia typing.
- This reduced panel offers a rapid and cost-effective method for immunophenotyping.
- The findings support immediate therapeutic decisions based on this simplified approach.
Background:
Immunophenotyping is at present a useful aid and often an essential step for correct diagnosis of acute leukemia and for this purpose some investigators have proposed several immunomarker panels. This approach is particularly important in the differential diagnosis of acute leukemias not classifiable by standard morphology and cytochemistry.
Methods:
We have tested peripheral blood and/or bone marrow samples from 125 patients not classifiable by FAB criteria. In all the cases, the reactivities of the same panel of 17 monoclonal antibodies were analyzed by flow cytometry, using both single and double fluorescent labeling.
Results:
Of the 125 patients investigated, 75 (60%) were classifiable as ALL, 58 as B-lineage ALL and 17 as T-lineage ALL; 33 (26.4%) as AML, of which 2 M7; 6 (4.8%) as biphenotypic and 11 (8.8%) as immunophenotypically undifferentiated.
Conclusions:
From a critical analysis of our cases and a review of the literature, we suggest that a panel of 9 monoclonal antibodies (CD2, CD5, CD7, CD10, CD19, CD20, CD13, CD33, CD41), is sufficient for reliable, rapid and reasonably low cost typing of acute leukemia, useful for an immediate therapeutic decision.
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