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A polystyrene biosensing chip to be used as a medical diagnostic tool has been developed. Its surface functionalization is optimized to bind the bioanalytes with high efficiency. Coatings of allyldextran monolayers were carried out on polystyrene chips activated with g-irradiation. The effect of the concentration of allyldextran solution on the coating efficiency was studied by surface-sensitive x-ray photoelectron spectroscopy (XPS) and contact-angle measurements. In a second step, sodium periodate chemistry was applied to functionalize the dextran layer, followed by coupling with streptavidin or neutravidin.

<b>Background:</b> The human platelet alloantigens (HPA) HPA-1a and HPA-5b are located on glycoproteins on the platelet surface and are the most relevant to cause neonatal alloimmune thrombocytopenia (NAIT). The antigens are defined by single nucleotide polymorphisms (SNPs) in the glycoprotein genes, and the antigen status can be determined by genotyping the SNPs. However, genotyping is time-consuming and costly depending on the method and sample throughput. Here, we tested the reliability of the evanescence wave based fluorescence (EVA) biosensor technology for the rapid phenotyping of the HPA-1a and HPA-5b antigens on blood donor samples in two laboratories. <b>Methods:</b> HPA-1a and HPA-5b phenotyping was performed on EDTA blood samples from 336 blood donors (Lyon: 216 donors; Mannheim: 120 donors) using EVA typing assays and the biosensor system (Davos Diagnostics, Davos, Switzerland). For genotyping, validated PCR-SSP and TaqMan-PCR methods were used. <b>Results:</b> HPA-1a phenotyping was positive for all samples with HPA-1aa (n = 244; EVA value 807 ± 167 U/s) and HPA-1ab (n = 82; 542 ± 110 U/s) genotypes. All samples (n = 10) with negative EVA values (<10 U/s) had the HPA-1bb genotype. HPA-5b phenotyping was negative for all HPA-5aa genotypes (n = 267) and positive for the HPA-5ab (n = 66; 83 ± 22 U/s) and HPA-5bb (n = 3; 118 ± 25 U/s) genotypes. EVA values from heterozygotes were significantly lower compared to HPA-1a or HPA-5b homozygotes. A strong correlation of the EVA values with the platelet count in the blood samples was observed. <b>Conclusion:</b> EVA is a reliable method for rapid phenotyping of the clinically relevant HPA-1a and HPA-5b platelet antigens. All phenotyping results were 100% concordant with the HPA-1 or HPA-5 genotype. The test can be performed from only 10 µl of fresh or frozen blood samples within less than 15 min time-to-result.