High-throughput multiplex HLA-typing by ligase detection reaction (LDR) and universal array (UA) approach.

One major goal of genetic research is to understand the role of genetic variation in living systems. In humans, by far the most common type of such variation involves differences in single DNA nucleotides, and is thus termed single nucleotide polymorphism (SNP). The need for improvement in throughput and reliability of traditional techniques makes it necessary to develop new technologies. Thus the past few years have witnessed an extraordinary surge of interest in DNA microarray technology. This new technology offers the first great hope for providing a systematic way to explore the genome. It permits a very rapid analysis of thousands genes for the purpose of gene discovery, sequencing, mapping, expression, and polymorphism detection. We generated a series of analytical tools to address the manufacturing, detection and data analysis components of a microarray experiment. In particular, we set up a universal array approach in combination with a PCR-LDR (polymerase chain reaction-ligation detection reaction) strategy for allele identification in the HLA gene.

[1]  S. P. Fodor,et al.  Determination of ancestral alleles for human single-nucleotide polymorphisms using high-density oligonucleotide arrays , 1999, Nature Genetics.

[2]  Andrea Frosini,et al.  Polymorphism analysis within the HLA‐A locus by universal oligonucleotide array , 2004, Human mutation.

[3]  Francis Barany,et al.  Universal DNA array detection of small insertions and deletions in BRCA1 and BRCA2 , 2000, Nature Biotechnology.

[4]  N C Dracopoli,et al.  Progress in high throughput SNP genotyping methods , 2002, The Pharmacogenomics Journal.

[5]  M. Colonna,et al.  HLA-A typing: comparison between serology, the amplification refractory mutation system with polymerase chain reaction and sequencing. , 1997, Tissue antigens.

[6]  N. Gerry,et al.  Universal DNA microarray method for multiplex detection of low abundance point mutations. , 1999, Journal of molecular biology.

[7]  Katarina Lindroos,et al.  Minisequencing on oligonucleotide microarrays: comparison of immobilisation chemistries , 2001, Nucleic Acids Res..

[8]  M. Alpers,et al.  High-Throughput SNP Genotyping , 2002, Comparative and functional genomics.

[9]  C. Consolandi,et al.  A structured chitosan-based platform for biomolecule attachment to solid surfaces: application to DNA microarray preparation. , 2006, Bioconjugate chemistry.

[10]  L Peltonen,et al.  Minisequencing: a specific tool for DNA analysis and diagnostics on oligonucleotide arrays. , 1997, Genome research.

[11]  Francis Barany,et al.  Multiplex PCR/LDR for detection of K-ras mutations in primary colon tumors , 1999, Oncogene.

[12]  O. Olerup,et al.  HLA-DR typing by PCR amplification with sequence-specific primers (PCR-SSP) in 2 hours: an alternative to serological DR typing in clinical practice including donor-recipient matching in cadaveric transplantation. , 1992, Tissue antigens.