Detection kit for TMEM101 gene methylation in human peripheral blood circulating tumor DNA for early screening of endometrial cancer

A technology of endometrial cancer and human peripheral blood, applied in the biological field, can solve the problems of untraceable blood early screening for endometrial cancer, long detection time, and high false positives, and achieve good patient compliance, easy operation, and easy interpretation simple effect

Active Publication Date: 2021-08-06
上海伯豪生物技术有限公司
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  • 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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  • Detection kit for TMEM101 gene methylation in human peripheral blood circulating tumor DNA for early screening of endometrial cancer
  • Detection kit for TMEM101 gene methylation in human peripheral blood circulating tumor DNA for early screening of endometrial cancer
  • Detection kit for TMEM101 gene methylation in human peripheral blood circulating tumor DNA for early screening of endometrial cancer

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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 the process of locking the candidate regions in this patent, the marker screening idea 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 thre...

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 non-methylated 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 prim...

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 detection kit for TMEM101 gene methylation in human peripheral blood circulating tumor DNA for early screening of endometrial cancer. The detection kit is applied to early screening of endometrial cancer and comprises a methylation specific primer of a TMEM101 gene target site, a hydrolysis probe which is specific to the TMEM101 gene, and a primer of a reference gene and a hydrolysis probe of the reference gene, wherein the target site of the TMEM101 gene is at least one CpG site in an upstream and downstream 2000bp interval of a transcription start site of the TMEM101 gene; the reference gene is one or more of GAPDH and beta-actin; and the kit comprises a PCR pre-amplification reaction system and a PCR amplification reaction system. According to the provided detection kit for TMEM101 gene methylation in human peripheral blood circulating tumor DNA for early screening of endometrial cancer, the technical defects that in the prior art, the process is tedious, the detection time is long, the false positive is high, and the early screening of the endometrial cancer blood cannot be traced are overcome.

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 Applications(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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