Molecular markers, internal reference genes and their applications, detection kits and methods for constructing detection models

A molecular marker, internal reference gene technology, applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc. Excessive treatment and inappropriate treatment, satisfying individualized precision treatment, and improving the effect of technical methods

Active Publication Date: 2021-06-29
CAPITALBIO CORP +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these products are currently developed based on European and American populations. After these products enter China, they are not only expensive, but whether the genes and their detection models are suitable for Chinese populations has yet to be verified.

Method used

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  • Molecular markers, internal reference genes and their applications, detection kits and methods for constructing detection models
  • Molecular markers, internal reference genes and their applications, detection kits and methods for constructing detection models
  • Molecular markers, internal reference genes and their applications, detection kits and methods for constructing detection models

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] Example 1 Collection of Samples, Sample Data Consolidation

[0063] The system collects complete clinical follow-up information without treatment of the initial breast cancer patient, and the inventors have selected 341 samples as TLDA (TAQman Lowdeysity Array, TLDA) chip detection. And subsequent series of QRT-PCR verification experimental samples:

[0064] (1) Unexpected early, medium-term breast cancer first diagnosis;

[0065] (2) Patients with Uneavenated early, medium ER or (and) PR positive breast cancer;

[0066] (3) LN or (and) no transfer, and the number of LN transfer;

[0067] (4) It has accurate and detailed follow-up information.

Embodiment 2

[0068] Example 2TLDA chip Screening molecular marker and interference gene

[0069] Two-six cases of breast cancer prognosis in compliance with the above conditions were detected by TLDA chip detection for TLDA chip detection. The specific steps are:

[0070] (1) Extraction of RNA in the FFPE sample: 8 tablets of 20 μm per sample 4 or 10 μm slices are taken, and RNA is extracted according to the highpure ffpet rna isolation kit (roche), and the extraction RNA is quantitatively controlled by NanoDrop-2000 After the downstream reverse transcription experiment.

[0071] (2) Total RNA is reactive to CDNA samples: Take 1μg total RNA according to Vilo TM The Master Mix Kit (Invitrogen) manual is reversed.

[0072] (3) CDNA sample for TLDA test: above CDNA products and After the Universal PCR Mastermix is ​​fully mixed, the test experiment was performed on the ABI 7900 fluorescent quantitative PCR instrument in accordance with the TLDA standard program.

[0073] (4) Data analysis and p...

Embodiment 3

[0077] Example 3 Molecular marker's large sample amount QRT-PCR verification

[0078] TLDA Screening 14 molecular markers and 5 innergic genes: BCl2, PGR, Scube2, ESR1, MKI67, CCNB1, MYBL2, GRB7, Erbb2, MMP11, CD68, BAG1, MAPT, MS4A1; ACTB, GAPDH, GUSB, NUP214 . The complete 289 FFPE samples in accordance with the above sample collection requirements and clinical follow-up information are used, and single tube QRT-PCR verification is performed.

[0079] (1) 289 cases of FFPE sample RNA extract: 8 μm slices per sample 4 or 10 μm sections are taken, and RNA is extracted according to the highpure ffpet RNA ISOLATION KIT (ROCHE). The extracted RNA is quantified by NanoDrop-2000 The downstream reverse transcription experiment is performed after the control.

[0080] (2) 289 cases of FFPE sample RNA reverse recorded cDNA: 1 μg total RNA according to Vilo TM Mastermix Kit (Invitrogen) Manual for reverse transcription.

[0081] (3) 289 cases of FFPE samples CDNA product for QPCR detectio...

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Abstract

The invention relates to the field of biotechnology, in particular to a primer composition and its application, a detection kit and a method for constructing a detection model. Using the follow-up information as a comparison method, the kit provided by the present invention has a prediction accuracy of 70% for the risk of recurrence or death in 3-10 years after the operation of newly diagnosed patients with ER and PR positive breast cancer, and the prediction accuracy of the low-risk group and the high-risk group The accuracies are 81.1% and 54.4%, respectively. The corresponding predictive accuracies of FFPE pathological test results were 71.9% and 56.8%, respectively. The risk prediction model matched with the kit only needs the molecular marker Ct value, patient age, pT stage, and LN number, without relying on other clinicopathological information. Reduce inappropriate treatment caused by pathological prediction errors, and further improve the technical method of breast cancer prognosis evaluation.

Description

Technical field [0001] The present invention relates to the field of biotechnology, and more particularly to molecular markers, internal paramers genes, and their applications, detection kits, and construction methods of detection models. Background technique [0002] Breast cancer is one of the main reasons why women's lives in women around the world. In 2013, the US Cancer Association issued the Normal Cancer Statistics showed that the incidence of breast cancer was first in women's cancer, and mortality was second. The latest US national cancer center data shows that in 2013, 232,340 cases of women have new breast cancer, 39,620 deaths. In the United States, there is an average of 8 women in the 8 women. Although China belongs to the low-issuing country with breast cancer, the incidence rate and mortality rate in recent years have increased significantly. About 15% from China in 10 million patients with newly diagnosed in the world. China's breast cancer network statistics sho...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6886C12N15/11
CPCC12N15/11C12Q1/68
Inventor 郭弘妍孙义民王亚辉谢展邢婉丽程京邓涛张治位
Owner CAPITALBIO CORP
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