[HLA-DQA1 genotyping by using oligonucleotide microarrays].
In order to fabricate the HLA-DQA1 genotyping chip and develop an integrated, parallel technical platform to type HLA system, a pair of primers and a set of probes were designed according to the sequences of HLA-DQA1 exon 2, where the polymorphism is concentrated. The oligonucleotide chip was made with the methods developed in our laboratory. The target DNA was asymmetrically amplified with the labeled sense primer. The signals were scanned and analyzed after the hybridization between microarray and PCR product. The allele types of the samples were identified. The result was verified by the standard DNA and DNA sequencing. The results showed that the genotyping was successfully carried out in 50 standard DNA samples and 50 clinical samples. Among them, results of the 50 standard DNA samples matched their templates. In the other 50 samples, results of the randomly selected 10 matched their sequencing results except that two of them got the incompletely result. In reproducible tests, the signal reappear rate was 95%. It is concluded that HLA-DQA1 genotyping by using our array system is simple and convenient with satisfied accuracy and reproducibility.
Department of Biochemistry and Molecular Biology, Shenyang Medical College, Shenyang 110034, China.
Tong
T
Wang
Tian-Jiao
TJ
Wang
Qun
Q
He
Yu-Kui
YK
Zhang
Jia-Ming
JM
Ma
Wei-Jian
WJ
Hou
Shao-Cheng
SC
Wang
Zhong-Cheng
ZC
Pan
Yu-Jie
YJ
Zhao
D005838:Genotype; D006683:HLA-DQ Antigens / Q000235:genetics* / Q000276:immunology; D059848:HLA-DQ alpha-Chains; D006801:Humans; D020411:Oligonucleotide Array Sequence Analysis; D015345:Oligonucleotide Probes; D020133:Reverse Transcriptase Polymerase Chain Reaction
D006683:HLA-DQ Antigens; D059848:HLA-DQ alpha-Chains; C069050:HLA-DQA1 antigen; D015345:Oligonucleotide Probes