Multicolor fluorescence PCR kit and method for detecting HLA-B*5801 allele

Abstract

The invention provides a multicolor fluorescence PCR kit and method for detecting HLA-B*5801 allele. The multicolor fluorescence PCR kit is characterized by comprising a detection primer-probe combination of two forward primers, two reverse primers and three detection probes for detecting the gene HLA-B*5801 and an internal-control primer probe comprising a forward primer, a reverse primer and an internal-control probe for detecting a reference gene. Based on the primer design and PCR reaction characteristics, the time and consumables can be saved to a great degree, the detection process only needs 1-1.5 hours, the operation is simple and easy to implement, and the whole experiment can be finished within 2.5 hours.

Description

technical field [0001] The invention relates to a multicolor fluorescent PCR kit for detecting HLA-B*5801 alleles, and also relates to a method for detecting HLA-B*5801 alleles, belonging to the field of gene diagnosis. Background technique [0002] Human leukocyte antigens (Humanleukocyte antigens, HLA) is the expression product of human major histocompatibility complex, in the immune system, it is mainly responsible for mutual recognition between cells, induction of immune response, and regulation of immune response. HLA is divided into three types: class I molecules are HLA-A, -B, -C series antigens, which are widely expressed on the surface of nucleated cells in various tissues; class II molecules are HLA-D / DR, -DP, DQ series antigens, mainly Expressed in B cells and antigen-presenting cells, class I and class II antigens are related to organ transplantation, of which class II antigens are more important; class III molecules are complement components. HLA is the gene co...

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Problems solved by technology

[0007] However, due to the sequence polymorphism of the HLA gene family, compared with the detection of HLA alleles, this technology has rarely been applied to the detection of HLA-B*5801 alleles. The fundamental reason is that multiple HLA-B alleles The gene and HLA-B*5801 have extremely high sequence homology (more than 90%), so a set of primer-probe combinations suitable for multicolor fluorescent PCR reactions to amplify HLA-B*5801 alleles with high specificity are designed very difficult

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