[Preliminary study on HLA-B genotyping by oligonucleotide chips].
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.
Lan
Ke
K
Shenzhen Yishengtang Biological Products Co. Ltd, Shenzhen 518026, China.
Hu
Shou-Wang
SW
Zhang
Fan
F
Wang
Hui
H
Guan
Wei
W
Ding
Yu
Y
Sun
Ou-Jun
OJ
Wang
Sheng-Qi
SQ
D005838:Genotype; D015235:HLA-B Antigens / Q000145:classification / Q000235:genetics*; D006801:Humans; D020411:Oligonucleotide Array Sequence Analysis; D017422:Sequence Analysis, DNA
D015235:HLA-B Antigens