Oligonucleotide microarray for HLA-DRB1 genotyping: preparation and clinical evaluation.
In this study, HLA-DRB1 gene was genotyped by using the microarray technique. Oligonucleotide probes were designed based on partial sequences of various genotypes of HLA-DRB1, and were fixed on a silylated slide to form a microarray. The second exon of HLA-DRB1 gene in the extracted genomic DNA samples was amplified and labelled by means of polymerase chain reaction (PCR); then it was hybridized to the microarray. The microarray was scanned, and the result was analysed in order to determine the genotypes of HLA-DRB1 of the tested sample. A total of 1574 of 1592 clinical samples had accordant results of genotypes in either microarray assay or PCR-SSP assay; 8 of 10 samples that had inconsistent results of genotypes were proved to be microarray-assay reliable by confirmation of DNA sequencing. It is concluded that microarray is an alternative reliable method for HLA-DRB1 genotyping.
Zhang
F
F
Beijing Institute of Radiation Medicine, Beijing 100850, China.
Hu
S W
SW
Huang
J
J
Wang
H
H
Zhang
W
W
Geng
Y Y
YY
Wang
S Q
SQ
D002681:China; D005091:Exons / Q000235:genetics; D005787:Gene Frequency; D005802:Genes, MHC Class II; D005838:Genotype; D006684:HLA-DR Antigens / Q000235:genetics*; D059811:HLA-DRB1 Chains; D006650:Histocompatibility Testing / Q000379:methods*; D006801:Humans; D020411:Oligonucleotide Array Sequence Analysis; D015345:Oligonucleotide Probes; D016133:Polymerase Chain Reaction / Q000379:methods; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013698:Templates, Genetic
D006684:HLA-DR Antigens; D059811:HLA-DRB1 Chains; D015345:Oligonucleotide Probes