[Clinical application for reference strand mediated conformation analysis system in HLA-A genotyping].
OBJECTIVE
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.


METHODS
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.


RESULTS
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 needed to be typed for 1 - 3 times to confirm their types.


CONCLUSIONS
RSCA has some advantages compared with the PCR-SSP typing method, which are high resolution, high sensitivity, high accuracy, high replication, capability to find new alleles, high throughput, low cost and suitability for unrelated hematopoietic stem cell transplant donor-recipient HLA typing. But there are still some defects in this new strategy, which are time-consuming for only one sample a time and higher DNA quality requirement. In addition, RSCA database needs to be further improved.
Zhao
Dan-dan
DD
Department of Immunology, Affiliated Hospital of Academy of Military Medical Sciences, Beijing 100039, China.
Sun
Yu-ying
YY
Xi
Yong-zhi
YZ
Jin
Li
L
Liu
Nan
N
Liang
Fei
F
D000293:Adolescent; D000328:Adult; D000368:Aged; D002648:Child; D005260:Female; D005838:Genotype; D015234:HLA-A Antigens / Q000235:genetics*; D006650:Histocompatibility Testing / Q000379:methods*; D006801:Humans; D008297:Male; D008875:Middle Aged; D018807:Polymorphism, Single-Stranded Conformational; D012680:Sensitivity and Specificity
D015234:HLA-A Antigens