Combined marker for endometrial benign and malignant lesions, detection primer probe set and kit

A technology for detection of benign and malignant diseases and primers, which is applied in the determination/testing of microorganisms, recombinant DNA technology, biochemical equipment and methods, etc., can solve the problems of non-invasive detection, achieve intuitive interpretation, increase accuracy, and reduce errors Effect

Inactive Publication Date: 2022-05-13
BEIJING ORIGIN-POLY BIO-TEC CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, there is no method for non-invasive detection of multiple uterine lesions such as uterine fibroids, uterine sarcoma, and endometrial cancer at the same time.

Method used

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  • Combined marker for endometrial benign and malignant lesions, detection primer probe set and kit
  • Combined marker for endometrial benign and malignant lesions, detection primer probe set and kit

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Experimental program
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Effect test

Embodiment 1

[0028] The Illumina Infinium Methylation EPIC BeadChip chip (including 853,307 CpG sites) was screened for methylated genes by selecting 20 cases of uterine fibroids, 20 cases of uterine sarcoma and 20 cases of endometrial cancer paraffin-embedded tissue samples.

[0029] Through the significant methylation difference site algorithm, the detection results of different types of endometrial benign and malignant samples were compared and analyzed, and the deltaBeta (positive methylation level of endometrial malignancy-benign endometrium control sample) was screened out. The 26 genes shared by the top 100 as gene markers are: ADCY4, ADRA1A, AMPD3, CDO1, CELF4, CHAD, CYP26C1, DRD4, FZD7, GNA12, GRM6, GYPC, HAND2, HSPA1L, HS3ST2, ITGA5, LTB4R2, MAGI2, MTMR7, NDUFS2, NEFM, PTGDR, RASSF1, SPDYA, TJP2, TRH. Methylation-specific PCR amplification verification was then performed. Because there are many CpG regions in each gene, it needs to be transferred to RT-PCR for verification and a...

Embodiment 2

[0032] Detection of endometrial benign and malignant lesion-related genes ADCY4, ADRA1A, AMPD3, CDO1, CELF4, CHAD, CYP26C1, DRD4, FZD7, GNA12, GRM6, GYPC, HAND2, HSPA1AL, HS3ST2, ITGA5, LTB4R2, MAGI2, MTMR7, NDUFS2, NEFM , PTGDR, RASSF1, SPDYA, TJP2, detection test of TRH methylation kit.

[0033] The specific nucleotide sequences of the primers and probes used are shown in the table below:

[0034] ADCY4-F1 GGGAGGGGGTCGCG

SEQ ID NO:1 ADCY4-R1 GAAATCACGCTCGCCGCTACG SEQ ID NO:2 ADCY4-FP1 FAM-GCGTTTGTCGTTCGGG-BHQ1 SEQ ID NO:3 ADRA1A-F1 TAGGTAGTTTTTCGGGATCG

SEQ ID NO:4 ADRA1A-R1 GACGCACTCACCTAAAACGCCG SEQ ID NO:5 ADRA1A-FP1 FAM-TTTCGCGCGCGAGTT-BHQ1 SEQ ID NO:6 AMPD3-F1 GTTGGGAAAAGTTGTGCG

SEQ ID NO:7 AMPD3-R1 CCGCCGAACTTCCACACCG SEQ ID NO:8 AMPD3-FP1 FAM-GACGTCGGGTAAGTTCGGGC-BHQ1 SEQ ID NO:9 CDO1-F1 ataac GTTTATATTTTTAAGTTATCG

SEQ ID NO:10 CDO1-R1 gattagACCCTCTACTAAT...

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Abstract

The invention discloses an endometrial benign and malignant lesion combined marker, a detection primer probe group and a kit, the combined marker comprises specific methylation regions of 26 target genes, detection primers and probes aiming at the methylation regions can be used for detection through fluorescent quantitative PCR, and the kit has the advantages of high sensitivity, high sensitivity and high sensitivity. A detection result adopts a logistic regression algorithm to evaluate the risk of benign and malignant endometrial lesions, including effective distinguishing of benign and malignant lesions such as hysteromyoma, hysterosarcoma, atypical endometrial hyperplasia and endometrial cancer. According to the invention, through multi-gene methylation combined detection, the sensitivity and specificity of detection of malignant endometrial lesions are significantly improved. The kit is particularly suitable for differential diagnosis of benign and malignant endometrial lesions by taking cervical exfoliated cells as samples, and has the advantages of simplicity and convenience in sampling, high detection speed, high accuracy, high specificity and the like.

Description

technical field [0001] The invention relates to a combined detection marker for endometrial benign and malignant lesions, a detection primer probe set and a kit. Background technique [0002] Uterine fibroids are the most common benign tumors in female genitalia, also known as fibroids, uterine fibroids. The incidence rate increases with age, and it is more common in women aged 30 to 55. Uterine fibroids can undergo degeneration, most of which are benign, including hyaline degeneration, cystic degeneration, and red degeneration, and a few become malignant, such as sarcomatoid. When malignant, it increases rapidly in a short period of time with vaginal bleeding, and does not shrink after menopause On the contrary, it becomes larger and is basically malignant. There are also some malignant transformations that are asymptomatic and easily overlooked. [0003] Since the development mechanisms of uterine sarcoma and endometrial cancer are different, an objective, repeatable, a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12Q1/686C12N15/11
CPCC12Q1/6886C12Q1/686C12Q2600/154C12Q2537/143C12Q2563/107
Inventor 王林海刘沛
Owner BEIJING ORIGIN-POLY BIO-TEC CO LTD
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