JMD TIDES - Oligonucleotide and Peptide - May 18-21, 2008, Las Vegas, NV
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JMD 2005, Vol. 7, No. 5
Copyright © 2005 American Society for Investigative Pathology & Association for Molecular Pathology

Simultaneous Genotyping of DRB1/3/4/5 Loci by Oligonucleotide Microarray

Ye Bang-Ce*, Chu Xiaohe{dagger}, Fan Ye{ddagger}, Li Songyang*, Yin Bincheng* and Zuo Peng*

From the State Key Laboratory of Bioreactor Engineering, * East China University of Science and Technology, Shanghai; Shenghua BioK Bio Co. Ltd, {dagger} Huzhou, Zhejiang; and Jiangnan Biotechnology Inc., {ddagger} Huzhou, Zhejiang, China

Matching of the HLA antigens for donor-recipient in transplantation, disease predisposition or protection, population studies, and forensic testing requires accurate but simple typing methods. Here, we describe a DNA-based tissue-typing assay that determines the haplotype of the DRB1/3/4/5 loci in hy-bridization of oligonucleotide array after sample amplification. Using this multianalyte DNA hybridization system, we analyzed seven regions of exon 2 of DRB loci that have single-base discrimination. Thirty-six oligonucleotide probes complementary to the alleles of interest were immobilized on each microslide. The efficiency and specificity of identifying DRB genotypes using the oligonucleotide arrays was evaluated by blinded analysis of 147 samples from reference standards and subjects. The established method provides a rapid and inexpensive DRB "low-resolution" typing tool for prescreening a large number of samples.




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