Preliminary study on HLA-B genotyping by oligonucleotide chips.
- Author:
Ke LAN
1
;
Shou-Wang HU
;
Fan ZHANG
;
Hui WANG
;
Wei GUAN
;
Yu DING
;
Ou-Jun SUN
;
Sheng-Qi WANG
Author Information
1. Shenzhen Yishengtang Biological Products Co. Ltd, Shenzhen 518026, China.
- Publication Type:Journal Article
- MeSH:
Genotype;
HLA-B Antigens;
classification;
genetics;
Humans;
Oligonucleotide Array Sequence Analysis;
Sequence Analysis, DNA
- From:
Journal of Experimental Hematology
2003;11(2):174-178
- CountryChina
- Language:Chinese
-
Abstract:
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.
