Data di Pubblicazione:
2004
Citazione:
Polymorphism analysis within the HLA-A locus by universal oligonucleotide array / C. Consolandi, A. Frosini, C. Pera, G.B. Ferrara, R. Bordoni, B. Castiglioni, E. Rizzi, A. Mezzelani, L. Rossi Bernardi, G. De Bellis, C. Battaglia. - In: HUMAN MUTATION. - ISSN 1059-7794. - 24:5(2004 Nov), pp. 428-434.
Abstract:
Human leukocyte antigen (HLA) class I genes present some of the most
complex single nucleotide polymorphism (SNP) patterns in the human genome.
HLA typing is therefore extremely challenging. In this article, we use the
ligation detection reaction (LDR) combined with a universal array (UA) as
a robust and efficient method to analyze SNPs within the HLA-A region that
includes HLA-A alleles of interest for immunotherapy in tumor diseases.
The LDR, combined with a UA platform, has been optimized for the detection
of 27 alleles distributed within exons 2 and 3 of HLA-A. The assay
involves the amplification by PCR of the HLA-A genomic region (1,900 bp),
the cycled ligation reaction, followed by the capture of ligated products
through hybridization onto a UA. Each slide was designed to allow the
detection of up to eight samples in parallel. The PCR/LDR/UA HLA-A assay
was evaluated by analyzing 62 individuals (31 homozygous and 31
heterozygous) previously typed by direct sequencing. We demonstrate that
the microarray genotyping procedure described here is a robust and
efficient method for unambiguous detection of HLA alleles. HLA genotyping
by PCR/LDR/UA is in perfect agreement with typing obtained by direct
sequencing. Our results clearly demonstrate that the combination of
enzymatic processing (LDR) and a demultiplexing hybridization onto a UA is
a robust tool for SNP discrimination within the highly polymorphic HLA
region. We demonstrate the specificity and efficiency of such an approach,
suggesting the feasibility of a PCR/LDR/UA low resolution HLA typing
procedure.
Tipologia IRIS:
01 - Articolo su periodico
Keywords:
DNA Mutational analysis methods, HLA-A antigens Genetics, Oligonucleotide Array Sequence analysis methods
Elenco autori:
C. Consolandi, A. Frosini, C. Pera, G.B. Ferrara, R. Bordoni, B. Castiglioni, E. Rizzi, A. Mezzelani, L. Rossi Bernardi, G. De Bellis, C. Battaglia
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