Rapid anti‐HPA‐1a antibody quantification with a Luminex bead‐based assay: A method evaluation

TL;DR

This study evaluates a new Luminex bead-based assay for quickly measuring anti-HPA-1a antibodies, which are linked to severe fetal and neonatal blood disorders.

Contribution

The study introduces a rapid and accurate alternative to the labor-intensive MAIPA assay for quantifying anti-HPA-1a antibodies.

Findings

The Luminex PakLx assay showed 97.6% agreement with the MAIPA assay in classifying results as positive or negative.

The correlation between PakLx MFI values and MAIPA IU/mL was strong (R² = 0.92).

PakLx enables faster laboratory processing and provides semi-quantitative results for clinical use.

Abstract

High concentration of human platelet antigen‐1a (HPA‐1a) antibodies is reported to be associated with severe foetal and neonatal alloimmune thrombocytopaenia (FNAIT). The gold standard for quantification of anti‐HPA‐1a antibodies is the monoclonal antibody immobilization of platelet antigen (MAIPA) assay, which is a laborious method performed in only a few reference laboratories. The aim of this study was to evaluate the performance of the commercially available bead‐based Luminex assay PakLx (Immucor) for quantitative measurement of anti‐HPA‐1a antibodies.

We analysed anti‐HPA‐1a antibody levels in plasma samples from 42 HPA‐1a‐negative women who had given birth to a child with thrombocytopaenia. Quantification of antibodies was performed with two different techniques: MAIPA analysed by spectrophotometry with results expressed in international units (IU)/mL, and PakLx analysed in the Luminex assay with results expressed as the mean fluorescence intensity (MFI).

In the comparison of the two methods' ability to stratify a result as either positive or negative, PakLx demonstrated 97.6% agreement with the MAIPA assay, with positive and negative predictive values of 96.7% and 100%, respectively. The correlation of the MFI values from PakLx with IU/mL in MAIPA assay was high, with a correlation coefficient (
R
2
) of 0.92. MFI values were converted into semi‐quantitative results: high, intermediate and low levels of anti‐HPA‐1a.

PakLx shows high agreement with the MAIPA assay and allows fast laboratory turnaround time for the determination of anti‐HPA‐1a antibody levels. The result may be of predictive value in clinical assessments.

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