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[Development of oligoneucleotide arrays for HLA-DR53-associated group genotyping]
Jia-quan Xiao1, Cheng-tao Li, Jian-ming Tan
1Department of Urology, Shanghai First People's Hospital, Shanghai Organ Transplant and Research Center, Shanghai 200080, China.
Objective:
To develop a method performed on an oligonucleotide array for HLA-DR53 group genotyping.
Methods:
According to the specific allelic frequency and sequence of HLA-DRB loci in Chinese Han population, HLA-DR53 group typing probes which were immobilized on a glass supports were synthesized. A pair of group-special primers labeled by the Cy5-dCTP were designed, and the primers were used in the PCR, thus the PCR products were labeled with Cy5. The labeled PCR products were hybridized with array. The signals were scanned by scanner and analyzed by image software. The typing results were confirmed by standard DNA and PCR-SSO. One hundred and eleven samples were typed by this array.
Results:
There were 72 HLA-DR53 group loci typed by oligonucleotide array. Among them, 34 loci were DR9, 25 were DR4, and 13 were DR7. No false positive or false negative typing results were observed. The specificity and reproducibility were 100% and the overall time of genotyping was 5 hours.
Conclusion:
The oligonucleotide array technique is a precise, rapid molecular method for HLA-DR53 genotyping, suited for clinical practice.