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

Zhao, Dan-dan; Sun, Yu-ying; Xi, Yong-zhi; Jin, Li; Liu, Nan; Liang, Fei.
Zhonghua Yi Xue Za Zhi; 83(8): 654-7, 2003 Apr 25.
Artículo en Chino | MEDLINE | ID: mdl-12887822

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.

CONCLUSION:

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.
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