11 research outputs found
New Triphosphate Conjugates Bearing Reporter Groups:Â Labeling of DNA Fragments for Microarray Analysis
CrossHub: a tool for multi-way analysis of The Cancer Genome Atlas (TCGA) in the context of gene expression regulation mechanisms
An RNA microchip containing immobilized oligoribonucleotides with protective groups at 2′-O- positions
Evaluation of a Low-Density Hydrogel Microarray Technique for Mycobacterial Species Identification
Plasmon-Induced Light Absorption of Phthalocyanine Layer in Hybrid Nanoparticles: Enhancement Factor and Effective Spectra
Analysis of T-Cell Receptor-γ Gene Rearrangements Using Oligonucleotide Microchip: A Novel Approach for the Determination of T-Cell Clonality
T-cell clonality estimation is important for the differential diagnosis between malignant and nonmalignant T-cell proliferation. Routinely used methods include polymerase chain reaction (PCR) analysis of T-cell receptor-γ (TCR-γ) gene rearrangements followed by Genescan analysis, polyacrylamide gel electrophoresis, or heteroduplex analysis to visualize amplification products. Here, we present a new method for the analysis after PCR of TCR-γ rearrangements using hybridization on oligonucleotide microchip. A microchip was designed to contain specific probes for all functional variable (V) and joining (J) gene segments involved in rearrangements of the TCR-γ locus. Fluorescently labeled fragments of rearranged γ-chain from patients and donors were obtained in a multiplex nested PCR and hybridized with a microchip. The results were detected using a portable microchip analyzer. Samples from 49 patients with T-cell lymphomas or leukemias and 47 donors were analyzed for T-cell clonality by microchip and single-strand conformation polymorphism analysis, which served as a standard reference method. Comparison of two techniques showed full concordance of the results. The microchip-based approach also allowed the identification of V and J gene segments involved in the particular TCR-γ rearrangement. The sensitivity of the method is sufficient to determine 10% of clonal cells in the sample