| Autor: |
Gail Flickinger, Romy Kronstein, Nezih Cereb, Soo Young Yang, HwaRan Kim, Amaralingeswara Rao, JeongOk Jeon, DongYong Kim, Torsten Tonn, Sang Yeol Seo, Jangyoung Kwon |
| Rok vydání: |
2016 |
| Předmět: |
|
| Zdroj: |
Human Immunology. 77:24 |
| ISSN: |
0198-8859 |
| DOI: |
10.1016/j.humimm.2016.07.041 |
| Popis: |
Aim One of the factors that needs to be considered for a successful hematopoietic stem cell transplantation is the ABO Rh compatibility between patient and donor. Until recently, most donor registries did not have ABO Rh typing data on donors due to logistics and expenses that are involved. Furthermore, most donor recruitments are currently done using buccal swabs or saliva samples that precludes serotyping for ABO Rh groups. However, these specimen types are suitable for DNA typing of ABO Rh genes. We have developed a method based on Illumina MiSeq platform to genotype ABO Rh genes and build an algorithm to predict the phonotypes. This additional information on donors could speed up the search process for the patients who need hematopoietic stem cell transplant. Methods: We designed 9 Exon-based primer pairs for the 7 exons for the glycosyltransferase gene. Exon 7 required 3 overlapping amplicons for full coverage. For Rh genes, we designed 10 primer sets to amplify all 10 exons. Previously, 1000 serotyped DNA samples of Caucasian ethnicity were genotyped to predict genotype-phenotype correlation. We applied a phenotyping algorithm based on all the exon sequences, incorporating known serological motif sequences in exons 6 and 7. This algorithm predicted ABO type with 99.9% accuracy and Rh with 100%. Results We blindly tested 1376 samples of diverse ethnic origin. We found that 12 (0.87%) samples for ABO and 31 (2.23%) samples for Rh had discordant results with serotyping. We typed close to 1 million individuals as of today, and we have discovered many novel sequences for each exon as shown in the table below: We have found 144 cases with new motifs in exon 7 that changed B blood group genotype to O. Thirteen had a point substitution, and 131 had a 1 bp deletion. Conclusion Large scale molecular typing for ABO and Rh is now feasible. Molecular basis for some ambiguous serotyped could be identified with this approach. ABO and Rh information in donor files will help accelerate the donor search process . |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
|
Full Text from ScienceDirect