Assessing reagents and techniques for identifying RhCE variants in routine serological testing

Area of Science:

• Transfusion Medicine
• Immunology
• Genetics

Background:

• RhCE variants are challenging to detect serologically, often identified only after antibody formation.
• Early identification of RhCE variants is critical for preventing Rh alloimmunization.
• Variable antigen expression complicates routine RhCE typing.

Purpose of the Study:

• To evaluate monoclonal antisera and various methodologies for detecting RhCE variants.
• To define reactivity patterns of RhCE variants with different expression levels.
• To assess the efficacy of different serological tests in identifying RhCE variants.

Main Methods:

• Samples with atypical RhCE reactivity were tested using tube, gel, and microplate methods with five antisera.
• Antigen expression was quantified by flow cytometry.
• RhCE variants were molecularly classified.

Main Results:

• Tube testing, especially with specific anti-e and anti-c clones, was most effective in identifying weak or partial RhCE antigens (63%-77%).
• Automated phenotyping and gel tests identified some variants but less effectively than tube tests for weak expression.
• Specific RhCE variants like RHCE*ceJAL were detected by automated methods.

Conclusions:

• The tube test plays a vital role in routine serological diagnostics for RhCE variants.
• Selecting appropriate monoclonal antibody clones is essential for variant detection.
• Early detection of reduced RhCE antigen expression aids molecular investigation and prevents alloimmunization.