Genotyping of HLA-DR1,DR51-associated group by DNA microarray / 中华泌尿外科杂志

ABSTRACT

Objective To develop a DNA microarray for HLA-DR1,DR51 group genotyping. Methods According to the specific allelic sequences coding HLA-DR1,DR51 loci,HLA- DR1,DR51 group typing probes which were immobilized on a glass support were synthesized.A pair of group-specific primers labeled by the Cy5-dCTP were designed,then the primers were used in the PCR,thus the PCR products were labeled with Cy5.The labeled PCR products were hybridized with array.The signals were scanned by scanner and analyzed by image software.The typing results were confirmed by standard DNA and PCR-SSO. Results A total of 130 samples were typed by this DNA array.There were 34 HLA-DR1,DR51 group loci typed by DNA array.Among them,18 loci were DR15,8 were DR16,6 were DR10 and 2 were DR1.No false positive or false negative typing results occurred.The accuracy and reproducibility were 100% and the overall time of genotyping was about 3.5 hours. Conclusions DNA array technique is a precise,rapid molecular method of high resolution power and high specificity for HLA-DR1,DR51 genotyping,which is applicable to clinical transplant practice.