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Kit for rapid detection of mRNA expression level of BRCA1 gene

A kit and expression technology, applied in the field of kits for rapid detection of BRCA1 gene mRNA expression, to achieve the effects of low false positive, high accuracy, and less pollution

Inactive Publication Date: 2012-01-11
李艳 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, there is no literature report on the kit for detecting the expression of BRCA1 gene mRNA

Method used

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  • Kit for rapid detection of mRNA expression level of BRCA1 gene
  • Kit for rapid detection of mRNA expression level of BRCA1 gene
  • Kit for rapid detection of mRNA expression level of BRCA1 gene

Examples

Experimental program
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Effect test

Embodiment 1

[0029] Embodiment 1. Preparation of kit of the present invention

[0030] (1) The kit of the present invention consists of the following:

[0031] ①Trizol: rapid extraction of cancer tissue RNA;

[0032] ② First strand cDNA synthesis kit (RT-PCR); including 25mmol / L MgCl 2 4μl, 10× reverse transcriptase buffer 2μl, 10mmol / L dNTP 2μl, RNase inhibitor 0.5μl, Oligo(dT) 15 0.5μg, AMV reverse transcriptase 15U, RNase-free deionized water;

[0033] ③ Primers: including target gene BRCA1, internal reference gene GAPDH and fluorescent probes, details are as follows:

[0034] BRCA1 gene primer sequence:

[0035] The upstream primer sequence of BRCA1 gene is: 5'-ATGGGCCCTTCACCAACA-3'; (SEQ ID No.1)

[0036] The downstream primer sequence of BRCA1 gene is: 5'-CACAGAAGCACCACACAGCTGTA-3'; (SEQID No.2)

[0037] Taqman fluorescent probe: 6FAM 5'-ATCAACTGGAATGGATG-3'TAMRA; (SEQ ID No.3)

[0038] GAPDH gene primer sequence:

[0039] The upstream primer sequence of GAPDH gene is: 5'-AT...

Embodiment 2

[0046] Example 2. The expression level of BRCA1 mRNA detected by the kit prepared in Example 1

[0047] Take the detection results of 20 cases of breast cancer paraffin section specimens as an example.

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Abstract

The invention relates to a fluorescence quantitative PCR (Polymerase Chain Reaction) diagnostic kit for rapid detection of mRNA expression level of a BRCA1 gene. The kit comprises BRCA1gene primer, a reference gene GAPDH primer and a Taqman fluorescence probe. The BRCA1 is an important cancer suppressor gene, and proteins coded by the BRCA1 gene play an important role in DNA damage and repair. A fluorescence quantitative PCR with high sensitivity and specificity is employed to detect the mRNA expression level of BRCA1; and a detection result has substantially increased specificity and sensitivity. The kit provides a novel rapid simple gene diagnostic technique for whether a clinical malignant tumor patient should use platinum chemotherapeutics and antimicrotubular medicaments.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a kit for rapidly detecting the expression level of BRCA1 gene mRNA by using a fluorescent quantitative PCR technique. Background technique [0002] Platinum compounds are the most commonly used tumor chemotherapy drugs, including cisplatin, carboplatin and oxaliplatin, etc., which have the characteristics of broad anticancer spectrum and strong effect. Because platinum-based drugs bind DNA extensively, causing cross-linking between DNA strands or intra-strand cross-linking, causing DNA damage, resulting in cell death to achieve the purpose of anti-cancer. Clinical application has shown that there are significant individual differences in the efficacy of platinum-based drugs. After platinum-based cytotoxic chemotherapy drugs damage tumors, the DNA repair mechanism of tumor cells is activated, and tumor cells with high repair ability are prone to resistance to chemotherapy. BRCA1 pla...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68G01N21/64
Inventor 李艳童永清
Owner 李艳
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