Kit for jointly detecting breast cancer 21 genes and preparation method of kit

A combined detection and kit technology, applied in the field of medical molecular biology, can solve the problems of error-prone, time-consuming, cumbersome operation process, etc.

Inactive Publication Date: 2014-08-27
杭州美中疾病基因研究院有限公司
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  • Abstract
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  • Application Information

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Problems solved by technology

However, the PCR reaction itself is a highly sensitive experiment, and so many genes are combined into reaction wells, wh

Method used

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  • Kit for jointly detecting breast cancer 21 genes and preparation method of kit
  • Kit for jointly detecting breast cancer 21 genes and preparation method of kit
  • Kit for jointly detecting breast cancer 21 genes and preparation method of kit

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Embodiment Construction

[0161] In order to make the purpose, technical solution and advantages of the present invention clearer, the present invention will be further described in detail below with reference to examples. It should be understood that the examples described here are only used to explain the present invention, and are not intended to limit the present invention.

[0162] The present invention adds known primers of 21 genes on the basis of common fluorescent quantitative PCR kit, and premixes 21 tubes of reaction solution. Each gene primer corresponds to the corresponding master mix tube and is marked. Then perform the premix performance test. Specific steps are as follows:

[0163] (1) extraction of volunteer breast cancer paraffin specimen mRNA, obtain sample 1, 2, 3 respectively: (using QIAGEN company FFPE Kit)

[0164] 1. First with a blade, trim the wax block.

[0165] 2. Cut into 5 μm wax slices on a paraffin microtome, and take 5-10 slices.

[0166] 3. Immediately put the t...

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Abstract

The invention discloses a kit for jointly detecting breast cancer 21 genes. A PCR (polymerase chain reaction) amplification primer consists of 21 primer pairs, and can be used for jointly detecting 21 gene-related expression quantities: Ki67, STK15, Survivin, CCNB1, MYBL2, MMP11, CTSL2, GRB7, HER2, GSTM1, CD68, BAG1, ER, PGR, BCL2, SCUBE2, ACTB, GAPDH, PRLPO, GUS, and TFRC. Output fragments of the 21 pairs of primers related in the invention are smaller than 100bp, so that PCR efficiency after reverse transcription of mRNA (messenger ribonucleic acid) extracted by a paraffin specimen is greatly improved, all primers of 21 genes are premixed, and time and labor for designing and synthesizing 21 pairs of genes by an experimenter are saved; the corresponding primers of the 21 genes are not needed to be added for premixing in mother liquor, and therefore, errors of reaction holes are greatly reduced. The kit disclosed by the invention is simple to operate, and capable of detecting on a quantitative PCR instrument by only adding a template, so that detection experimental time is greatly shortened. The kit disclosed by the invention is applicable to a single sample and multiple samplers, so that results can be quickly obtained, and therefore, scientific research and clinical detection requirements can be satisfied very well.

Description

technical field [0001] The invention belongs to the field of medical molecular biology and relates to a kit, in particular to a kit for combined detection of breast cancer 21 genes. Background technique [0002] In recent years, with the in-depth study of genomics, the "21 gene detection" (Oncotype Dx) for estrogen receptor positive patients has been proved to be more accurate than clinicopathological indicators in predicting estrogen receptor positive, axillary lymph node Negative and reduce the risk of distant metastasis in patients taking tamoxifen. The test is the only multigene test recommended jointly by the American Society of Clinical Oncology (ASCO) and National Comprehensive Cancer Network (NCCN) guidelines. The 21 genes included 16 tumor-related genes and 5 reference genes. Tumor-related genes include: proliferation-related genes (Ki-67, STK15, Survivin, Cyclin B1, MYBL2), invasion-related genes (Stromelysin3, Cathepsin L2), Her-2-related genes (GRB7, Her-2), ho...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6886C12Q2600/158
Inventor 陈伟
Owner 杭州美中疾病基因研究院有限公司
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