Method for evaluating curative effect of medication to cure HIV through detecting change of TCR gene of human T cell receptor before and after treatment
A cell receptor and gene technology, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve problems such as low sensitivity and specificity, and achieve highly reproducible results
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[0029] Elaborate the specific implementation mode of the present invention below in conjunction with embodiment:
[0030] Blood samples from patients receiving AIDS drug treatment and healthy volunteers were collected separately, separated with Fico11-Hypaque layering fluid, and after horizontal centrifugation, mononuclear cells were densely concentrated in the interface between the plasma layer and the layering fluid, and the cells in this layer were aspirated After being washed and resuspended, PBMCs were obtained. CD4 or CD8 T cells were further isolated with magnetized beads coupled with CD4 or CD8 antibody.
[0031] The method for isolating and purifying the mRNA of T cells is to adopt the published technology (Chen, Z.W et al. 1994 proc, Matl. Acai. Sci. Usa. 91: 7501-7505), mRNA with 1 μg arbitrary hexanucleotides (Promega, the United States Madison, Wisconsin), 5 μl supertranscriptase II (Gibco BRL, Long Island, New York, USA) with a total volume of 20 μl, mixed at 37...
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