Clinical application for reference strand mediated conformation analysis system in HLA-A genotyping

To establish and stabilize a new HLA typing strategy, reference strand mediated conformation analysis (RSCA), and to conform its advantages by using HLA-A as target gene. 20 standard DNA samples from international RSCA cooperative team were used to establish and stabilize RSCA. 84 DNA samples of related hematopoietic stem cell transplant donor-recipients were extracted from peripheral blood cells. HLA-A loci were typed both by RSCA and PCR-SSP. RSCA results of 20 standard samples were identical to those of international RSCA cooperative team, and the accuracy and replication rate were both 100%. Among the 84 samples, 82/84 (97.6%) cases could be designated definitely, and 33/84 (39%) cases could be typed to allelic level. In addition, 2/84 (2.4%) cases could not be detected by RSCA for their poor PCR results. 20 samples were randomly selected to identify the replication rate of RSCA, and the results demonstrated that the replication rate was 100%. Among the PCR-SSP typing results, there were about 10% samples