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Kit for detecting 15 gene mutation sites related to rectal cancer chemoradiotherapy sensitivity and use thereof

A mutation site, radiotherapy and chemotherapy technology, applied in the field of biomedical diagnosis, can solve the problems of limited efficiency and difficult labeling, and achieve the effect of avoiding complicated detection and saving time and labor costs.

Active Publication Date: 2021-06-18
广州瑞熹生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The effectiveness of current clinical and molecular indicators for predicting the efficacy of neoadjuvant chemoradiotherapy is very limited, limited to the selection of research objects based on theory and experience, and it is doomed to be difficult to efficiently mine a large number of sub-markers that affect the outcome of the study

Method used

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  • Kit for detecting 15 gene mutation sites related to rectal cancer chemoradiotherapy sensitivity and use thereof
  • Kit for detecting 15 gene mutation sites related to rectal cancer chemoradiotherapy sensitivity and use thereof
  • Kit for detecting 15 gene mutation sites related to rectal cancer chemoradiotherapy sensitivity and use thereof

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Embodiment 1

[0030] The diagnostic kit for predicting neoadjuvant chemoradiotherapy sensitivity of locally advanced rectal cancer based on gene DNA mutation site of the present invention, said diagnostic kit for predicting neoadjuvant chemoradiotherapy sensitivity of locally advanced rectal cancer based on gene DNA mutation site includes Specific primers for detecting gene mutation sites, the nucleotide sequences of the upstream primers of the specific primers are as shown in SEQ ID NO.1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21 , 23, 25, 27, 29, the downstream primer nucleotide sequence of the specific primer is as SEQ ID NO.2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22 , 24, 26, 28, 30.

[0031]所述的基因DNA突变位点如下:chr6_33052958_C_T、 chr9_125316157_C_T、chr9_123850770_G_A、chr3_122259606_T_C、 chr1_150679033_G_A、chr6_160211646_GTT_、chr1_160920966_C_T、 chr8_23294761_T_C、chr21_47974582_A_G、chr7_122635024_C_T、chr6_56420538_C_T、chr1_232574921_T_C、chr8_21862551_A_G、 chr11_66254085_G_T、chr11_44636833_G_A。

[0032]...

Embodiment 2

[0045] Collection and preparation of tumor tissue samples

[0046] The inventor collected tumor tissue samples of locally advanced rectal cancer from September 2005 to June 2017, and these groups met the inclusion criteria. The inclusion criteria mainly include four points: 1. Locally advanced rectal patients diagnosed for the first time and who have not received any treatment; 2. Patients receiving standard treatment of neoadjuvant chemoradiotherapy, radical surgery and postoperative adjuvant chemotherapy; Confirmed by examination, TGR grading score, PCR group is TRG 1, that is: pathological complete response (pathological complete response, PCR) after neoadjuvant chemotherapy, and none-PCR group is TRG 2-5, that is: neoadjuvant chemotherapy Group with residual tumor (None-PCR group); 4. Those who are willing to participate in the study and cooperate with follow-up. According to the principle of gender and age matching, the samples of radiochemotherapy sensitive group and ra...

Embodiment 3

[0054] Whole Exome Sequencing and Data Analysis

[0055] The inventors selected tumor tissue samples from 8 neoadjuvant chemoradiotherapy-sensitive patients and 7 neoadjuvant chemoradiotherapy-insensitive patients for whole exome sequencing screening. There were no significant differences in gender, mean age, and distribution of these patients (Table 1). The clinicopathological characteristics of the specimens used for exome analysis are shown in Table 1:

[0056] The present invention adopts the whole exome sequencing method of Themor Fisher Company to screen differential gene mutation sites between neoadjuvant chemoradiotherapy-sensitive patients and neoadjuvant chemoradiotherapy-insensitive patients, and exome sequencing can detect the exon regions of all protein-coding genes Mutation frequency of DNA mutation sites. See the Themor Fisher website for specific steps. After calibration of the obtained raw data, the inventors used the Ion Reporter analysis method of the Ion...

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Abstract

The invention discloses a kit for detecting 15 gene mutation sites related to rectal cancer chemoradiotherapy sensitivity and use thereof. The detection kit is based on a high-throughput DNA sequencing technology and used for detecting DNA mutation sites of 15 genes related to the rectal cancer chemoradiotherapy sensitivity, and comprises specific primers for detecting the gene mutation sites. The upstream primers of the specific primers have nucleotide sequences shown as SEQ ID NO.1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27 and 29 and the downstream primers of the specific primers have nucleotide sequences shown as SEQ ID NO.2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28 and 30. The kit has good stability for detecting gene DNA mutation sites of a tumor tissue specimen, high clinical operability and a simple and convenient detection process. The kit can be used for distinguishing a patient with local advanced rectal cancer sensitive to neoadjuvant radiotherapy and chemotherapy, such that the patient with local advanced rectal cancer is remarkably benefited in the neoadjuvant radiotherapy and chemotherapy.

Description

technical field [0001] The invention relates to the technical field of biomedical diagnosis, in particular to a kit for detecting mutation sites of 15 genes related to sensitivity to radiotherapy and chemotherapy of rectal cancer and its application. Background technique [0002] Among malignant tumors, the incidence of colorectal cancer ranks 3rd in the world and 5th in my country. Its incidence is rapidly rising and younger. It is a common malignant tumor that affects the development of our society and seriously endangers human health. Neoadjuvant chemoradiotherapy is the main component of the overall treatment strategy for rectal cancer, and the sensitivity of neoadjuvant chemoradiotherapy varies significantly among individuals. The degree of tumor regression after neoadjuvant chemoradiotherapy is significantly related to the long-term survival rate and quality of life of patients. About 30% of patients can achieve complete pathological remission during surgery after neoa...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12N15/11G16B20/50
CPCC12Q1/6886G16B20/50C12Q2600/156C12Q2600/106
Inventor 王穗海李明肖巍魏高远红
Owner 广州瑞熹生物科技有限公司
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