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Microarrays for high-throughput genotyping of MICA alleles using allele-specific primer extension.

Authors
Type
Published Article
Journal
Tissue Antigens
1399-0039
Publisher
Wiley Blackwell (Blackwell Publishing)
Publication Date
Volume
82
Issue
4
Pages
259–268
Identifiers
DOI: 10.1111/tan.12201
PMID: 24461005
Source
Medline
Keywords
License
Unknown

Abstract

The role of major histocompatibility complex (MHC) class I chain-related gene A (MICA), a ligand of NKG2D, has been defined in human diseases by its allele associations with various autoimmune diseases, hematopoietic stem cell transplantation (HSCT) and cancer. This study describes a practical system to develop MICA genotyping by allele-specific primer extension (ASPE) on microarrays. From the results of 20 control primers, strict and reliable cut-off values of more than 30,000 mean fluorescence intensity (MFI) as positive and less than 3000 MFI as negative, were applied to select high-quality specific extension primers. Among 55 allele-specific primers, 44 primers could be initially selected as optimal primer. Through adjusting the length, six primers were improved. The other failed five primers were corrected by refractory modification. MICA genotypes by ASPE on microarrays showed the same results as those by nucleotide sequencing. On the basis of these results, ASPE on microarrays may provide high-throughput genotyping for MICA alleles for population studies, disease-gene associations and HSCT.

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