Simultaneous genotyping of DRB1/3/4/5 loci by oligonucleotide microarray.
Matching of the HLA antigens for donor-recipient in transplantation, disease predisposition or protection, population studies, and forensic testing requires accurate but simple typing methods. Here, we describe a DNA-based tissue-typing assay that determines the haplotype of the DRB1/3/4/5 loci in hy-bridization of oligonucleotide array after sample amplification. Using this multianalyte DNA hybridization system, we analyzed seven regions of exon 2 of DRB loci that have single-base discrimination. Thirty-six oligonucleotide probes complementary to the alleles of interest were immobilized on each microslide. The efficiency and specificity of identifying DRB genotypes using the oligonucleotide arrays was evaluated by blinded analysis of 147 samples from reference standards and subjects. The established method provides a rapid and inexpensive DRB "low-resolution" typing tool for prescreening a large number of samples.
Bang-Ce
Ye
Y
State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China. bcye@ecust.edu.cn
Xiaohe
Chu
C
Ye
Fan
F
Songyang
Li
L
Bincheng
Yin
Y
Peng
Zuo
Z
D002460:Cell Line; D015342:DNA Probes; D005091:Exons / Q000235:genetics; D005456:Fluorescent Dyes; D005820:Genetic Testing / Q000379:methods*; D005838:Genotype; D006684:HLA-DR Antigens / Q000235:genetics*; D059811:HLA-DRB1 Chains; D006801:Humans; D007438:Introns / Q000235:genetics; D009693:Nucleic Acid Hybridization; D020411:Oligonucleotide Array Sequence Analysis; D016133:Polymerase Chain Reaction; D015203:Reproducibility of Results
D015342:DNA Probes; D005456:Fluorescent Dyes; D006684:HLA-DR Antigens; D059811:HLA-DRB1 Chains