Comparison of two high‐throughput platforms for red blood cell and platelet antigens genotyping

Abstract

Background Red blood cells (RBC) and platelet (PLT) antigens genotyping is a useful tool applied to solve complex immunohaematologic cases, perform alloimmunization prophylaxis and search for rare blood donors. There are different high‐throughput platforms available for RBC and PLT genotyping, which differ in terms of comprised antigens, length of reaction, result interpretation and cost. The objective of this study was to compare the performance of two high‐throughput platforms designed for RBC and PLT genotyping: OpenArray (customized Real‐time platform) and BLOODchip (Luminex technology). Materials and Methods Blood samples of 242 blood donors were simultaneously tested using OpenArray and BLOODchip (ID‐Core XT and ID‐HPA XT) platforms. The methodologies were compared in terms of accuracy, duration of reaction, percentage of no‐calls, number of samples processed per reaction and interpretation software. The designed OpenArray assay comprised 37 single nucleotide polymorphisms (SNPs) encoding 31 RBC antigens and six PLT antigens, while the BLOODchip comprised 29 SNPs encoding erythrocyte antigens (ID ‐Core XT) and 12 SNPs encoding PLT antigens (ID‐HPA XT). Results The OpenArray accuracy was 99·93%, and BLOODchip accuracy, 100%. The percentage of no‐calls did not differ between the methodologies. OpenArray was less time‐consuming when performing both RBC and PLT genotyping, whereas BLOODchip presented proper software for results interpretation and allowed the evaluation of individuals samples per reaction. Conclusion Both OpenArray and BLOODchip methodologies are accurate and suitable for RBC and PLT genotyping. The choice between the platforms relies on the service genotyping goals.

Implementing a fully automated high‐throughput strategy for blood donor genotyping

“…The great advantage of those arrays, besides their high accuracy, is the availability of its own software for results interpretation [4,9]. Open DNA-array platforms enable assay customization in accordance with the blood bank's needs, respecting the population heterogeneity of RBC and PTL antigens [4,8].…”

“…The SNPs selection was based on previous standardization experiments performed by our team . The OpenArray ® assay used in the present study was previously validated by our team by testing in parallel 242 samples with this methodology and with ID BLOODchip (ID Core XT and ID‐HPA XT .…”

“…The SNPs selection was based on previous standardization experiments performed by our team . The OpenArray ® assay used in the present study was previously validated by our team by testing in parallel 242 samples with this methodology and with ID BLOODchip (ID Core XT and ID‐HPA XT . In order to validate the SNPs that differed between the two platforms, Sanger sequencing was performed in 1% of the samples in the standardization step and confirmed allelic zygosity for Wra/Wrb, Kna/Knb and Sla/Vil.…”

“…DNA‐array strategies are much more attractive, as they can comprise, in a single reaction, the most clinically relevant RBC and PTL antigens . There are some array options specifically designed for blood donor genotyping: ID CORE XT (Progenika Biopharma‐Grifols, Bizkaia, Spain), Array Imaging System (AIS) with HEA BeadChip™ (BioArray Solutions, Immucor, Warren, NJ, USA), MassARRAY with Hemo ID Panels (Agena Bioscience Inc., San Diego, CA, USA), SNaPshot on ABI Capillary Electrophoresis Analyzer (Applied Biosystems, Foster City, CA, USA), MLPA SALSA (MRC‐Holland, Amsterdam, the Netherlands) and other open platforms that can be customized, like Real‐time PCR OpenArray (Life Technologies, Grand Island, NY, USA) . Any chosen DNA‐array methodology can become time‐consuming and prone to clerical mistakes if not associated with automated nucleic acid (DNA) extraction and polymerase chain reaction (PCR), as well as with complete data interfacing.…”

Effectiveness of a red cell antigen‐matching transfusion protocol in sickle cell disease patients

“…Donors were genotyped for the alleles RHCE*E/e, RHCE*C/c, KEL*1/2, FY*A/B, FY*B (-67T>C), JK*A/B and GYPB*S/s using a customized hybridization plate comprising 32 SNPs designed for the OpenArray â platform [18]. The assay consisted of primers and probes conjugated with VIC and FAM fluorophores, and each probe was designed to bind specifically to one allele type.…”