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Personalized Peptide Arrays for Detection of HLA Alloantibodies in Organ Transplantation
Published on: September 6, 2017
[DNA typing of HLA-I and II antigens with medium resolution method by DNA chip technique]
Gang Guo1, Rui Zhang, Ming-xin Zhang
1Tianjin Institute of Endocrinology, Tianjin 300052, China.
Objective:
To establish a DNA typing method for the HLA-I and II antigens in the Chinese of Han Nationality in Northern China with medium resolution method by DNA chip technique.
Methods:
Corresponding primers for the sense exon fragments of HLA-A, HLA-B, HLA-DR, HLA-DQB, and HLA-DQA were designed. A DNA typing chip was made with specific medium resolution typing probes designed according to the gene frequency of HLA-I and II alleles among the Chinese of Han nationality in Northern China. Unsymmetrical PCR was used to amplify the HLA-I and II exon2 and exon3 in 30 samples from the Chinese of Han nationality in Northern China, and then the PCR products were labeled and hybridized with the probes on the chip. The typing of HLA-I and II was certified by scanning the hybridized products and analyzing them with a set of computer software.
Results:
HLA-I and II alleles were successfully typed in 30 clinical samples. The h probes with medium resolution were able to discern 42 HLA-I and II alleles accurately. By using this chip 30 HLA-I and II alleles were distinguished and 6 new specific alleles were found.
Conclusion:
DNA typing of HLA-I and II by chip has been proven to be a high-resolution and high-specificity method. It is able to check out new alleles that can not be distinguished by other methods with the same resolution, and it is more intuitive and more suitable for clinical application.
