Microarray‐based genotyping for blood groups: comparison of gene array and 5′‐nuclease assay techniques with human platelet antigen as a model

Human platelet alloantigens (HPAs) and their corresponding alloantibodies (alloabs) play an essential role in fetal and neonatal alloimmune thrombocytopenia, posttransfusion purpura and platelet transfusion refractoriness. Emerging genotyping methods (e.g. microarray technology) will allow an automated throughput for HPA typing of large donor cohorts in the near future. For the clinical diagnosis of alloimmune thrombocytopenia, however, the proof of pathogenic alloabs still remains a prerequisite. Nowadays, monoclonal antibody-based antigen capture assays are the state of the art in many laboratories. This technique seems to come up against limiting factors. In a certain number of cases that are highly suspicious for alloimmune thrombocytopenia, platelet-specific alloabs are not detectable. Current observations indicate that these alloabs are heterogeneous in terms of their alloantigenic determinants as well as the consequences on platelet function. This heterogeneity may correlate to the severe bleeding complications sometimes seen in patients with alloimmune thrombocytopenia. New insights on the role of pathogenic alloabs will help us to improve diagnostic and therapeutic approaches for the adequate treatment of affected patients.

Section: Zusammenfassungmentioning

confidence: 99%

Heterogeneity of Platelet Alloantigens and Alloantibodies: New Insights into Structure and Function

Socher

Kroll

2006

“…In the method used by our group, the universal primers are labeled with a fluorochrome, and the final PCR product can be directly applied to the DNA arrays [9]. Also, in the method described by Bugert et al [10], the oligonucleotides used in the multiplex PCR are carrying a Cy- Red Blood Cell and Platelet Genotyping 263 3 label, and therefore the PCR product can be immediately analyzed by the DNA array. In the published papers, both Bugert et al [10] and our group [9] focus on the typing for all clinically relevant HPA genotypes -HPA-1, -2, -3, -5, -15 -as a proof of principle of glass array-based genotyping by allelespecific probe hybridization.…”

Section: High-throughput Genotyping Of Blood Donorsmentioning

confidence: 99%

“…Overall, 3 different types of genotyping methods were presented, which are illustrated in figure 3A, B, C. Our group and Bugert et al [9,10] used a similar method of genotyping by allele-specific hybridization to allele-specific probes spotted onto glass slides ( fig. 3A).…”

Section: High-throughput Genotyping Of Blood Donorsmentioning

confidence: 99%

“…Also, in the method described by Bugert et al [10], the oligonucleotides used in the multiplex PCR are carrying a Cy- Red Blood Cell and Platelet Genotyping 263 3 label, and therefore the PCR product can be immediately analyzed by the DNA array. In the published papers, both Bugert et al [10] and our group [9] focus on the typing for all clinically relevant HPA genotypes -HPA-1, -2, -3, -5, -15 -as a proof of principle of glass array-based genotyping by allelespecific probe hybridization. DNA microarray-based typing showed complete concordance with serological typing results or previously obtained typing results.…”

Section: High-throughput Genotyping Of Blood Donorsmentioning

confidence: 99%

See 1 more Smart Citation

Red Blood Cell and Platelet Genotyping: from Current Practice to Future High-Throughput Donor Typing

Haas

Schoot

Beiboer

et al. 2006

The molecular basis of almost all red cell and platelet blood group antigens is known. This enables the prediction of red cell or platelet phenotypes based upon the genotypes. In many laboratories, blood group genotyping assays are routinely used in cases where patient red cells cannot be used for serological typing due to the presence of autoantibodies (direct antiglobulin test (DAT)-positive) or after recent transfusion. Furthermore, fetal genotyping is routinely performed where there is a risk of alloimmune-mediated red cell or platelet destruction. In the Netherlands, medium-throughput human platelet antigen (HPA) genotyping has been introduced in the blood banks to guarantee direct (on-shelf) availability of HPA-1a- and HPA-5b-negative platelets for newborns suffering from neonatal alloimmune thrombocytopenia. The techniques used, pyrosequencing and Taq- Man-based allele-specific hybridization assay, are discussed in this review. Recently, several research groups investigated whether red cell molecular typing could also be introduced for large-scale blood donor typing to obtain and maintain an inventory of typed (antigen-negative) donors. The reason for this is that phenotyping of large cohorts of donors is a labor-intensive exercise and hampered by the lack of sufficient amounts of approved typing reagents for all blood group systems of interest. Several approaches for high-throughput genotyping, based on DNA microarrays spotted on glass slides or beads, have now been tested and are also discussed in this review.

“…managed by transfusion of matched platelets [49]. Assays like microarray-based genotyping or flow cytometric cross-match can be used for this purpose [50,51].…”

Section: Platelet Function Testing In Patients With Bleeding Disordersmentioning

confidence: 99%

State of the Art in Platelet Function Testing

Kehrel

Brodde

2013

Platelets perform many functions in hemostasis but also in other areas of physiology and pathology. Therefore, it is obvious that many different function tests have been developed, each one conceived and standardized for a special purpose. This review will summarize the different fields in which platelet function testing is currently in use; diagnostics of patients with bleeding disorders, monitoring patients' response to anti-platelet therapy, monitoring in transfusion medicine (blood donors, platelet concentrates, and after transfusion), and monitoring in perioperative medicine to predict bleeding tendency. The second part of the review outlines different methods for platelet function testing, spanning bleeding time, and platelet counting as well as determining platelet adhesion, platelet secretion, platelet aggregation, platelet morphology, platelet signal transduction, platelet procoagulant activity, platelet apoptosis, platelet proteomics, and molecular biology.