Construction of microarrays for genotyping of DQA using unmodified 45-mer oligonucleotide.
The human leukocyte antigen (HLA) class II system is strongly connected to immunological response and its compatibility between tissues is critical in transplantation. The simple robust typing analyses of HLA genes are extremely important. In this paper, we developed an approach based on microarray technology for genotyping of DQA gene. The microarrays were constructed with a total 31 unmodified 45-mer oligonucleotide. The second exon of DQA gene was amplified, and allowed to hybridize with the array. DQA genotypes were assigned by quantitative analysis of the hybridization results. The arrays were evaluated by DQA genotyping of nine reference samples and 120 clinical samples. The results demonstrate that the genotyping accuracy/concordance achieved 97.5% compared with the direct DNA sequencing. Although our methods did not perform high-resolution genotyping, it could be an alternative for serological typing in routine medical practice.
Yin
Bin-Cheng
BC
State Key Laboratory of Bioreactor Engineering, Department of Food Engineering and Science, East China University of Science & Technology, Shanghai 200237, China.
Yue
Fei
F
Ye
Bang-Ce
BC
D015342:DNA Probes / Q000737:chemistry*; D005838:Genotype; D006683:HLA-DQ Antigens / Q000235:genetics*; D006801:Humans; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D009841:Oligonucleotides / Q000737:chemistry*
D015342:DNA Probes; D006683:HLA-DQ Antigens; D009841:Oligonucleotides