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Related Experiment Videos

Assessing immunophenotyping performance: proficiency-validation for adopting improved flow cytometry methods.

Lance E Hultin1, Frederick A Menendez, Patricia M Hultin

  • 1Department of Medicine, Cellular Immunology and Cytometry, University of California, Los Angeles, California, USA.

Cytometry. Part B, Clinical Cytometry
|January 6, 2007
PubMed
Summary

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Implementing new flow cytometry methods for T-cell analysis can be streamlined. Evaluating performance using proficiency testing panels offers an alternative to repeated "switch" studies, accelerating adoption of superior techniques.

Area of Science:

  • Immunology
  • Laboratory Science
  • Clinical Diagnostics

Background:

  • Advancing immune cell phenotyping necessitates regular updates to laboratory methods and technology.
  • National Institute of Allergy and Infectious Diseases (NIAID)-sponsored research protocols require flow cytometry laboratories to conduct "switch" studies to validate new T-cell subset analysis methods.

Purpose of the Study:

  • To evaluate the impact of adopting a new 3-color, lyse no-wash flow cytometry method in two Multicenter AIDS Cohort Study (MACS) laboratories.
  • To assess alternatives to repeating failed "switch" studies for method validation.

Main Methods:

  • Comparison of a 2-color, lyse-wash method with a 3-color, lyse no-wash method across four MACS flow cytometry laboratories.
  • Evaluation of performance using the Immunology Quality Assurance (IQA) proficiency-testing program for laboratories that adopted the new method after failing initial switch studies.

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Main Results:

  • Laboratory performance at the two sites significantly improved after adopting the new method, as evidenced by IQA standard test results.
  • Reduced variation was observed across the four MACS sites and among replicate samples with the new method.

Conclusions:

  • Alternative validation strategies to repeating failed "switch" studies should be considered for adopting new laboratory methods.
  • Testing the new method against a proficiency testing reference panel or demonstrating superior performance on reference samples with both old and new methods are viable alternatives.
  • These alternatives can expedite the transition to superior methodologies, bypassing the need for multiple, serial "switch" studies.