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Use of IgM monoclonal reagents licensed for tube tests in column agglutination technology
F Morelati1, A Burlini, K J Reis
1Centro Trasfusionale e di Immunologia dei Trapianti, Istituto di Ricovero e Cura a Carattere Scientifico (IRCCS), Ospedale Maggiore, Milan, Italy.
Insights
Monoclonal IgM reagents licensed for tube tests show valid results with column agglutination technology (CAT) for red blood cell (RBC) phenotyping. This study confirms their suitability for broader application in blood typing.
Area of Science:
- Transfusion Medicine
- Immunology
- Hematology
Background:
- Red blood cell (RBC) phenotyping is crucial for safe transfusions.
- Current column agglutination technology (CAT) has limitations with certain reagents.
- Monoclonal IgM reagents are widely used in traditional tube tests.
Purpose of the Study:
- To evaluate the efficacy of monoclonal IgM reagents in column agglutination technology (CAT).
- To determine if reagents licensed for tube tests perform reliably with CAT.
- To assess the compatibility and performance of these reagents in a new platform.
Main Methods:
- Commercial CAT system was utilized.
- Two procedures (A and B) with varying reagent and RBC volumes were tested.
- Reagent suitability was assessed through inhibition tests, potency titrations, and specificity determinations.
- 1644 samples were typed comparing CAT with standard tube methods.
Main Results:
- All tested reagents allowed free passage of antigen-negative RBCs in CAT.
- Immediate-spin CAT methods yielded results consistent with tube methods.
- High agreement rates were observed: complete for Kell, 99.9% for Kidd, and 98.9-99.4% for Lewis reagents.
- Discrepancies were minimal, primarily with Lewis reagents in specific patient populations.
Conclusions:
- Monoclonal IgM reagents licensed for tube tests are largely suitable for use with CAT.
- CAT offers a viable alternative for RBC phenotyping using these established reagents.
- Further validation may be needed for specific reagents like Lewis, especially in complex cases.
Background:
Red cell (RBC) phenotyping using column agglutination technology (CAT) is currently limited by the reagents formulated in the system. To overcome this limitation, it was investigated whether monoclonal IgM reagents licensed for use with tube tests produced valid results with CAT.
Study Design And Methods:
Commercial CAT, does not contain antisera, was used to evaluate Procedures A (40 microL of reagent and 10 microL of 4% RBCs) and B (50 microL of reagent and 50 microL of 0.8% RBCs) with or without incubation at room temperature. In Study 1, reagents were tested to determine whether potentiators inhibit the passage of antigen-negative RBCs through the column. In Study 2, CAT sensitivity was measured by the use of potency titrations to define a procedure for each reagent that matched or exceeded that of the tube method. In Study 3, the specificity of each reagent was determined in parallel with the CAT and tube tests. Typing of 1644 samples was performed.
Results:
Study 1: Free passage was obtained with all reagents. Study 2: Immediate-spin methods using CAT produced the same results as the tube method. Study 3: With 8048 comparisons made, discrepant results were found in 32 transfused patients and in 6 cord blood samples, mainly with Lewis reagents. With comparison of CAT and the standard tube method, complete agreement was obtained with Kell reagents, 99.9-percent agreement with Kidd reagents, and 98.9-percent and 99.4-percent agreement with Lewis reagents.
Conclusion:
Most examined reagents seem suitable for use with CAT.
