Kit for detecting ERCC1 mRNA (Excision Repair Cross Complement Group 1 Messenger Ribonucleic Acid) expression by using fluorescence quantitative PCR (Polymerase Chain Reaction) technology

A technology for technical detection and fluorescence quantification, applied in the field of kits for detecting ERCC1 mRNA expression, to achieve the effects of good specificity, low false positives, and high degree of automation

Inactive Publication Date: 2011-08-17
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] At present, there is no literature report on the kit for detecting the expression of ERCC1 mRNA

Method used

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  • Kit for detecting ERCC1 mRNA (Excision Repair Cross Complement Group 1 Messenger Ribonucleic Acid) expression by using fluorescence quantitative PCR (Polymerase Chain Reaction) technology
  • Kit for detecting ERCC1 mRNA (Excision Repair Cross Complement Group 1 Messenger Ribonucleic Acid) expression by using fluorescence quantitative PCR (Polymerase Chain Reaction) technology
  • Kit for detecting ERCC1 mRNA (Excision Repair Cross Complement Group 1 Messenger Ribonucleic Acid) expression by using fluorescence quantitative PCR (Polymerase Chain Reaction) technology

Examples

Experimental program
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Embodiment 1

[0037] Example 1. Preparation of the kit of the present invention

[0038] (1) test kit of the present invention is composed as follows:

[0039] ①Trizol: rapid extraction of cancer tissue RNA;

[0040] ②First-strand cDNA synthesis kit (RT-PCR);

[0041] ③ Primers: including the target gene ERCC1, the internal reference gene GAPDH and fluorescent probes, as follows:

[0042] ERCC1 gene primer sequence:

[0043] Upstream: 5′-GGGAATTTGGCGACGTAATTC-3′;

[0044] Downstream: 5′-GCGGAGGCTGAGGAACAG-3′;

[0045] Taqman fluorescent probe: 6FAM 5'-CACAGGTGCTCTGGCCCAGCACATA-3'TAMRA

[0046] β-actin gene primer sequence:

[0047] Upstream: 5′-TGAGCGCGGCTACAGCTT-3′;

[0048] Downstream: 5′-TCCTTAATGTCACGCACGATTT-3′;

[0049] Taqman fluorescent probe: 6FAM5'-ACCACCACGGCCGAGCGG-3'TAMRA.

[0050] The above primer sequences and probe sequences were synthesized by Shanghai Sangon Bioengineering Technology Service Co., Ltd.

[0051] ④Negative control and positive control: take deionized ...

Embodiment 2

[0054] Example 2. Detection of ERCC1 mRNA expression with the kit prepared in Example 1

[0055] Take the detection of the tissue results of 30 cases of non-small cell lung cancer paraffin section specimens as an example.

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Abstract

The invention relates to the field of biotechnology, discloses a fluorescence quantitative PCR (Polymerase Chain Reaction) diagnostic kit for rapidly detecting ERCC1 mRNA (Excision Repair Cross Complement Group 1 Messenger Ribonucleic Acid), and aims to provide a kit capable of rapidly, conveniently, sensitively and specifically detecting ERCC1 mRNA and application thereof. An expression status of a DNA (Deoxyribonucleic Acid) repair gene can be used as a relatively ideal predictor for a chemotherapeutic effect. Nucleotide excision repair (NER) is in close relation with platinum drug resistance, wherein the excision repair cross complement group 1 (ERCC1) is an important factor which plays a role in a repair process and causes platinum drug resistance; the ERCC1 mRNA level is detected by using higher-sensitivity and higher-specificity fluorescence quantitative PCR; and the specificity and the sensitivity of a detection result are obviously improved. The kit provides a completely new rapid and convenient gene diagnosis technology for use of platinum type chemotherapeutic medicaments of clinical malignant tumor patients.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a kit for detecting the expression level of ERCC1 mRNA by using fluorescence quantitative PCR technology. Background technique [0002] Since cisplatin was found to have anticancer activity in 1967, the application and research of platinum metal anticancer drugs have developed rapidly. Cisplatin has the characteristics of broad anti-cancer spectrum and strong effect. It is one of the most commonly used drugs in current chemotherapy. Platinum drugs bind to intracellular DNA, resulting in DNA inter-strand cross-chain or intra-strand cross-chain, causing DNA damage. resulting in cell death. However, after cytotoxic chemotherapy drugs such as cisplatin damage tumors, the DNA repair mechanism of tumor cells is activated, and tumor cells with high repair ability are more likely to be resistant to chemotherapy. [0003] Studies suggest that the expression of DNA repair genes can be used a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68G01N21/64
Inventor 刘辉刘亚莉张敬磊周伟平
Owner SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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