Simple image analysis for reliability control of column agglutination test in pretransfusion testing

Area of Science:

• Transfusion Medicine
• Clinical Laboratory Science
• Analytical Chemistry

Background:

• Indirect antihuman globulin test (IAT) is crucial for pretransfusion testing to detect erythrocyte alloantibodies.
• Failure to detect alloantibodies can lead to transfusion complications.
• Column agglutination technique (CAT)-based IAT relies on precise liquid separation by air gaps, which can be disrupted by pipetting errors.

Purpose of the Study:

• To develop a rapid, meniscus-based analysis for identifying analytical errors in CAT-based IAT.
• To investigate the fluid dynamics of CAT-based IAT and detect air gap penetration caused by pipetting errors.

Main Methods:

• Investigated fluid dynamics of CAT-based IAT.
• Developed an image-based quality control approach using routinely generated analyser images.
• Quantified meniscus contact angle using ImageJ software in 132 simulated crossmatch tests.

Main Results:

• Receiver operating characteristic analysis showed high discriminatory performance (AUC 0.935).
• Experiments confirmed that increased protein concentration significantly reduces meniscus contact angle.
• The meniscus contact angle reliably predicts pipetting errors.

Conclusions:

• The novel meniscus-based analysis offers a rapid method to identify unrecognised pipetting errors.
• This approach has the potential to enhance test reliability and improve patient safety in routine IAT CAT workflows.
• Further validation across platforms and laboratories is needed.