TMEM101 Gene Methylation Detection Kit for Early Screening of Endometrial Cancer in Human Peripheral Blood Circulating Tumor DNA

A technology for endometrial cancer and human peripheral blood, applied in biochemical equipment and methods, microbial determination/inspection, instruments, etc., can solve the problem of untraceable blood early screening for endometrial cancer, long detection time, and cumbersome process and other problems, to achieve the effect of good patient compliance, easy operation and simple interpretation

Active Publication Date: 2021-10-01
上海伯豪生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0014] The technical problem to be solved by the present invention is to overcome the technical deficiencies in the prior art, such as cumbersome process, long detection time, high false positives, untraceable blood early screening of endometrial cancer, etc., and provide a method for early screening of endometrial cancer. Kit for detection of TMEM101 gene methylation in human peripheral blood circulating tumor DNA

Method used

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  • TMEM101 Gene Methylation Detection Kit for Early Screening of Endometrial Cancer in Human Peripheral Blood Circulating Tumor DNA
  • TMEM101 Gene Methylation Detection Kit for Early Screening of Endometrial Cancer in Human Peripheral Blood Circulating Tumor DNA
  • TMEM101 Gene Methylation Detection Kit for Early Screening of Endometrial Cancer in Human Peripheral Blood Circulating Tumor DNA

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Effect test

Embodiment 1

[0070] The difficulty of the present invention is to find the site for distinguishing endometrial cancer from other tumors. In order to lock the site for distinguishing endometrial cancer and other tumors proposed in the present invention, more than 33 tumors in the database were used in the research and development process. The methylation numbers of nearly 13,000 tumors have been developed, and the analysis process has been developed by itself, and a large amount of tumor tissue data has been actually detected by pyrosequencing. Some results are displayed as figure 1 As shown, specifically:

[0071] 1. In this patent, in the process of locking candidate regions, the idea of ​​marker screening is divided into two parts: 1. Training set, 2. Test set

[0072] 1. Training set

[0073] 1.1 Using the 450K chip methylation data of 284 cases of endometrial cancer (2 / 3) and 46 cases of adjacent cancers in TCGA, after data preprocessing, difference analysis was performed, and the th...

Embodiment 2

[0089] Materials: Plasma samples to be tested, positive quality control products, and negative quality control products. The positive quality control product is fully methylated human genomic DNA modified by bisulfite, and the negative quality control product is unmethylated human genomic DNA modified by bisulfite.

[0090] TMEM101 gene methylation detection kit in human peripheral blood circulating tumor DNA, including: TMEM101 gene target site methylation-specific primers; TMEM101 gene-specific hydrolysis probes; internal reference gene primers and internal reference gene hydrolysis probes ;

[0091] The target site of the TMEM101 gene is at least one CpG site in the interval of 2000 bp upstream and downstream of the transcription initiation site of TMEM101; the internal reference gene is one or more of GAPDH and β-actin.

[0092] The methylation-specific primers for the target site of the TMEM101 gene (the final concentration is: 0.1-1 µM) include:

[0093] Forward primer...

Embodiment 3

[0133] Through the cfDNA detection of 20 cases of healthy controls and 26 samples of Shanghai xx Hospital (refer to Example 1 for the detection method), the detection sensitivity of the kit of the present invention is 87%, the specificity is 100%, and the accuracy rate is 96%. See Table 5 and Table 6.

[0134] Table 5 Comparison of clinicopathological information and experimental results of 26 samples from xx Hospital in Shanghai.

[0135]

[0136] Table 6 The detection results of the regions involved in the present invention in 46 samples.

[0137]

[0138] As can be seen from Table 5 and Table 6, the detection kit of the present invention has reliable detection results, high sensitivity, good specificity, high accuracy, simple interpretation, and can trace the source of endometrial cancer tissue, reaching the level of the prior art. Unforeseen technical effects.

[0139] In summary, the kits or reagents of the embodiments of the present invention have the following a...

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Abstract

The invention discloses a kit for detecting methylation of TMEM101 gene in human peripheral blood circulating tumor DNA for early screening of endometrial cancer, which is applied to early screening of endometrial cancer. The kit includes: TMEM101 gene purpose The methylation-specific primer of the site; the hydrolysis probe specific to the TMEM101 gene; the primer of the internal reference gene and the hydrolysis probe of the internal reference gene; the target site of the TMEM101 gene is in the interval of 2000 bp upstream and downstream of the transcription start site of the TMEM101 gene at least one CpG site; the internal reference gene is one or more of GAPDH and β-actin; the kit includes a PCR pre-amplification reaction system and a PCR amplification reaction system. The present invention overcomes the technical deficiencies in the prior art, such as cumbersome process, long detection time, high false positives, untraceable source of blood for early screening of endometrial cancer, etc., and provides a human peripheral blood circulation system for early screening of endometrial cancer. A kit for the detection of TMEM101 gene methylation in tumor DNA.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a detection kit for methylation of TMEM101 gene in circulating tumor DNA in human peripheral blood for early screening of endometrial cancer. Background technique [0002] Endometrial cancer is a common malignant tumor of the female reproductive tract. With the increase of the average life expectancy of the population and the change of living habits, the incidence of the disease has shown a trend of continuous increase and rejuvenation in the past 20 years. In Western countries, the incidence of endometrial cancer has occupied the first place among malignant tumors of the female reproductive system. In my country, as the second most common gynecological malignancy after cervical cancer, its incidence accounts for about 20% to 30% of gynecological malignancies, and the incidence of endometrial cancer in some developed cities has reached the first place in gynecological ma...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6886C12Q1/6858G16B40/00
CPCC12Q1/6858C12Q1/6886C12Q2600/154C12Q2600/166G16B40/00C12Q2531/113C12Q2545/101C12Q2545/113C12Q2523/125
Inventor 王德韬孟宪欣徐惠敏李琎肖华胜
Owner 上海伯豪生物技术有限公司
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