Preliminary study on HLA-B genotyping by oligonucleotide chips / 中国实验血液学杂志

HLA genes constitute a highly polymorphic multigene system. In the present study, HLA-B oligonucleotide chips were manufactured by using a set of sequence-specific oligonucleotide probes derived from polymorphic regions in exon 2 and exon 3 of HLA-B gene spotted by microarrayer onto the aldehyde modified glass slides. In addition, the sequenced HLA-B gene clones used as standard samples were amplified from exon 2 and exon 3 by PCR. Together with the correct hybridization and wash conditions, the PCR products were bound with the array probes on the chip, and the hybridization patterns were transformed to HLA-B genotypes. The results showed that the genotypes of standard samples by the HLA-B oligonucleotide chips were completely identical with the sequenced clones. In conclusion, the oligonucleotide chip method presented here for HLA-B genotyping is a rapid, accurate, sensitive and attractive high throughput biochemical way.

HLA-A genotyping by oligonucleotide arrays / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate HLA genotyping by oligonucleotide arrays.</p><p><b>METHOD</b>Unsymmetrical PCR was used to amplify HLA-A gene exon 2, 3. The PCR products were used as templates for hybridization. The oligonucleotide probes were spotted on glass to make microarrays. High signal and specific probes were selected. The effects of the length and location of probes on hybridization signal were studied. A set of computer software was designed for scanning and genotype differentiation.</p><p><b>RESULT</b>The genotypes of 30 samples analyzed by microarray showed concordance to that by SBT and PCR-SSP.</p><p><b>CONCLUSION</b>HLA-A genotyping by oligonucleotide array is a good method with advantage of high speed, low cost and high flux.</p>