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Kit for detecting PDL-1 mRNA gene expression quantity and application thereof

A technology for PDL-1 and gene expression level, applied in the field of kits for detecting PDL-1 mRNA gene expression level, can solve the problems of inability to accurately reflect the expression level and expression level, the interpretation is greatly influenced by subjective factors, and the detection time is long. Achieve the effect of low detection cost, easy standardization, simple and fast operation

Inactive Publication Date: 2018-05-29
宿迁市第一人民医院
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  • Abstract
  • Description
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  • Application Information

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Problems solved by technology

[0004] At present, the methods used to detect the expression level of PDL-1mRNA mainly include immunohistochemical method, which takes a long time to detect, and the interpretation is greatly affected by subjective factors, and cannot accurately reflect the expression level and expression amount.

Method used

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  • Kit for detecting PDL-1 mRNA gene expression quantity and application thereof
  • Kit for detecting PDL-1 mRNA gene expression quantity and application thereof
  • Kit for detecting PDL-1 mRNA gene expression quantity and application thereof

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Embodiment Construction

[0021] The present invention also provides a method for detecting the gene expression of human PDL-1 mRNA based on the above kit, which specifically includes the following steps:

[0022] (1) Total RNA is extracted from peripheral blood, tumor tissue, puncture specimen or other specimens to be tested, and the concentration of the obtained RNA is measured.

[0023] (2) Perform reverse transcription operation on the obtained RNA sample to obtain cDNA;

[0024] Primer configuration: Oligo DT 0.5μl, Radom primer 0.5μl;

[0025] Add samples according to the following system: primer 1 μl, RNA template 10 μl,

[0026] RT-mix configuration: 5× reverse transcription buffer 4μl, dNTP 2μl, RNase Inhibitor 0.5μl, reverse transcriptase 1μl, DEPC water 1.5μl;

[0027] After the above samples were bathed in water at 65°C for 5 minutes, immediately place them on ice for 2 minutes; then add 9 μl RT-mix, centrifuge briefly in a centrifuge for 2-3 sec after adding, and place at room temperatur...

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Abstract

The invention relates to a kit for detecting a PDL-1 mRNA gene expression quantity. The kit mainly comprises specific primers and a PCR reaction reagent, and is characterized in that the sequences ofthe specific primers are as shown in the description; a PDL-1 upstream primer is as shown in the SEQ ID NO.1 and is 5'-AAGCACACGTGCCAAAGAATC-3'; and a PDL-1 downstream primer is as shown in the SEQ IDNO.2 and is 5'-TCTCGACGACGACGTTTCC-3'. The method can easily, quickly, accurately and reliably detect the gene expression quantity of the PDL-1 mRNA.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and in particular relates to a kit for detecting the expression level of PDL-1 mRNA gene and its application. Background technique [0002] Programmed death-1-ligand 1 (PD-L1) is an indispensable co-stimulatory molecule for T cell activation, widely expressed in activated T cells, B cells, macrophages and other cells . PD-1 / PD-L co-stimulatory signals are activated or inhibited depending on the degree of inflammation, immune status and genetic background of the body, and are mainly involved in the central and peripheral immune tolerance of T cells. The PD-L1 signaling pathway (as shown in the figure) plays a very extensive and complex role in immune regulation, and is closely related to autoimmune diseases, tumors, chronic infection and inflammation. A large number of studies have shown that after PD-L1 binds to the receptor PD1 on the surface of activated T cells, it can inhibit the ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6851C12Q1/6886
CPCC12Q1/6851C12Q1/6886C12Q2600/118C12Q2600/158C12Q2531/113C12Q2545/114
Inventor 裴兵
Owner 宿迁市第一人民医院
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