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Composition and kit for methylation noninvasive early detection of gynecological tumours

A technology for early detection of gynecological tumors, applied in recombinant DNA technology, biochemical equipment and methods, and microbial determination/examination. And other issues

Inactive Publication Date: 2021-06-08
BEIJING ORIGIN-POLY BIO-TEC CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the incidence of ovarian cancer is lower than that of the other two gynecological tumors, the mortality rate should not be underestimated, and the prognosis of patients with advanced ovarian cancer is often poor. According to data, about 75% of ovarian cancer patients in my country are advanced when they are first diagnosed, and they survive for 5 years less than 30%
However, the amount of tumor circulating DNA in plasma is small and prone to degradation

Method used

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  • Composition and kit for methylation noninvasive early detection of gynecological tumours
  • Composition and kit for methylation noninvasive early detection of gynecological tumours
  • Composition and kit for methylation noninvasive early detection of gynecological tumours

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0085] Detection kit for detection of methylation of gynecological malignancy-related genes PTGER4, HAND2, PAX1, HOXA9, FOXD3, OPCML, HS3ST2 and JAM3.

[0086] The specific nucleotide sequences of the primers and probes used are shown in the table below:

[0087]

[0088]

[0089] Note: F represents the forward detection primer, R represents the reverse detection primer, and FP represents the detection probe. In this table, the probe sequences shown have been fluorescently labeled and quenched.

[0090] The kit components are as follows:

[0091]

[0092]

[0093] Select 70 cases of ovarian cancer samples with known clear pathological information: 30 cases were identified as high-grade serous carcinoma, 20 cases of low-grade serous carcinoma, 10 cases of clear cell carcinoma, 10 cases of mucinous carcinoma; 40 cases were identified as benign ovarian samples .

[0094] Select 60 cases of endometrial cancer samples with known clear pathological information: 15 cas...

Embodiment 2

[0126] The specific nucleotide sequences of the primers and probes used are shown in the table below:

[0127]

[0128]

[0129] Note: F represents the forward detection primer, R represents the reverse detection primer, and FP represents the detection probe. In this table, the probe sequences shown have been fluorescently labeled and quenched.

[0130] Other components in the kit are the same as in Example 1.

[0131] Compared with clinicopathological results, the combined ROC curve area obtained by using this methylation detection kit is 0.894 (see Figure 4 ), the overall specificity was 97.5%, the detection sensitivity of ovarian cancer was 90%, the detection sensitivity of endometrial cancer was 86.7%, and the detection sensitivity of cervical cancer was 86%. Among them, the detection rates of high-grade serous ovarian cancer, low-grade serous ovarian cancer, clear cell ovarian cancer and mucinous ovarian cancer were 90%, 90%, 90% and 90% respectively; The detect...

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Abstract

The invention discloses a composition and kit for methylation noninvasive early detection of gynecological tumours. The composition comprises a target gene detection primer used for detecting methylation sites of target genes; and the target genes are PTGER4, HAND2, PAX1, HOXA9, FOXD3, OPCML, HS3ST2 and JAM3. The kit comprises a detection primer and a probe designed for the gene of the composition, a reference gene detection primer and a probe, a PCR reaction liquid and other reagents. By detecting the methylation state of the gene, noninvasive early screening can be simultaneously performed on various gynecological tumours at one time; and the method has the advantages of high detection speed, low cross contamination, final result accuracy ensured by a logistic regression algorithm and the like, and helps to realize the purposes of early discovery, early diagnosis and early treatment of malignant gynecological tumours.

Description

technical field [0001] The invention relates to the technical field of nucleic acid in vitro diagnosis, in particular to the application of specific gene methylation markers to the early detection and auxiliary diagnosis of gynecological tumors. Background technique [0002] Cervical cancer, endometrial cancer and ovarian cancer are the three most common malignant tumors in gynecological tumors in my country. The incidence of cervical cancer in my country is relatively high, and the incidence of endometrial cancer is also increasing year by year in recent years. As far as the survival rate of patients is concerned, the tumor cells of endometrial cancer are not easy to spread because they are confined to the uterus, and the 5-year survival rate of early stage patients can reach 70% to 80%. The 5-year survival rate can also reach about 80%. Although the incidence of ovarian cancer is lower than that of the other two gynecological tumors, the mortality rate should not be unde...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886C12Q1/6858C12N15/11
CPCC12Q1/6858C12Q1/6886C12Q2600/154C12Q2600/16C12Q2600/178C12Q2531/113C12Q2561/101C12Q2563/107C12Q2537/143C12Q2521/101
Inventor 刘沛王林海王紫芸
Owner BEIJING ORIGIN-POLY BIO-TEC CO LTD
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