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CN-122012711-A - Composition for in vitro detection of colorectal cancer and application thereof

CN122012711ACN 122012711 ACN122012711 ACN 122012711ACN-122012711-A

Abstract

The application provides a composition for detecting colorectal cancer and application thereof, wherein the composition comprises nucleic acid for detecting methylation state of a target gene, and the target gene is selected from one or more than two of MSC gene, FOXE gene, GDF6 gene and C9orf50 gene. The application also provides a kit comprising the composition and the use of the composition in the preparation of a kit for in vitro detection of colorectal cancer.

Inventors

  • Liu Shuanping
  • WANG XIUXIU
  • GUO YUANYUAN
  • DU BAOCHEN
  • WU ZHEN

Assignees

  • 博尔诚（北京）科技有限公司

Dates

Publication Date
20260512
Application Date
20260209

Claims (10)

  1. 1. A composition for detecting colorectal cancer in vitro, the composition comprising: nucleic acid for detecting methylation status of a target gene, Wherein the methylation state of the target gene is characterized by methylation of a target sequence of the target gene, Wherein the target gene is selected from one or more than two of MSC gene, FOXE gene, GDF6 gene and C9orf50 gene.
  2. 2. The composition according to claim 1, wherein the target sequence of the MSC gene is or comprises the sequence set forth in any one of SEQ ID NOs 1-4, and/or The target sequence of FOXE gene is the sequence shown in any one of SEQ ID NOs 5-8 or contains the sequence shown in any one of SEQ ID NOs 5-8, and/or The target sequence of the GDF6 gene is or comprises the sequence shown in any one of SEQ ID NOs 9-12, and/or The target sequence of the C9orf50 gene is a sequence shown in any one of SEQ ID NOs 13-16 or comprises a sequence shown in any one of SEQ ID NOs 13-16.
  3. 3. The composition of claim 1 or 2, wherein the nucleic acid for detecting methylation status of a target gene comprises: A primer which is a fragment of at least 9 nucleotides in a target sequence of the target gene, The fragment comprises at least one CpG dinucleotide sequence.
  4. 4. The composition of any one of claims 1-3, wherein the nucleic acid for detecting methylation status of a target gene comprises: A probe which is a fragment of at least 15 nucleotides hybridised to the target sequence of the target gene under moderately stringent or stringent conditions, The fragment comprises at least one CpG dinucleotide sequence.
  5. 5. The composition of any one of claims 1-4, further comprising: an agent that converts an unmethylated cytosine base at position 5 of a target sequence of a target gene into uracil.
  6. 6. The composition according to claim 3, wherein, The fragment of at least 9 nucleotides, which is the sequence shown as SEQ ID NO. 17 and SEQ ID NO. 18, or which is the sequence shown as SEQ ID NO. 20 and SEQ ID NO. 21, or which is the sequence shown as SEQ ID NO. 23 and SEQ ID NO. 24, or which is the sequence shown as SEQ ID NO. 26 and SEQ ID NO. 27.
  7. 7. The composition of claim 4, wherein the fragment of at least 15 nucleotides is the sequence set forth in SEQ ID NO. 19, or is the sequence set forth in SEQ ID NO. 22, or is the sequence set forth in SEQ ID NO. 25, or is the sequence set forth in SEQ ID NO. 28.
  8. 8. An oligonucleotide for detecting colorectal cancer in vitro, comprising: a fragment of at least 9 nucleotides of SEQ ID NO. 1 or SEQ ID NO. 2 or SEQ ID NO. 3 or SEQ ID NO. 4 or the complement thereof and comprising at least one CpG dinucleotide sequence, or A fragment of at least 9 nucleotides of SEQ ID No. 5 or SEQ ID No. 6 or SEQ ID No. 7 or SEQ ID No. 8 or the complement thereof and comprising at least one CpG dinucleotide sequence, or SEQ ID NO. 9 or 10 or 11 or 12 or a fragment of at least 9 nucleotides of the complementary sequence comprising at least one CpG dinucleotide sequence, or Fragments of at least 9 nucleotides of SEQ ID NO. 13 or SEQ ID NO. 14 or SEQ ID NO. 15 or SEQ ID NO. 16 or the complement thereof and comprising at least one CpG dinucleotide sequence.
  9. 9. The oligonucleotide of claim 8, further comprising: A fragment which hybridizes under moderately stringent or stringent conditions to at least 15 nucleotides of said SEQ ID NO. 1 or SEQ ID NO. 2 or SEQ ID NO. 3 or SEQ ID NO. 4 or the complement thereof and which comprises at least one CpG dinucleotide sequence; A fragment which hybridizes under moderately stringent or stringent conditions to at least 15 nucleotides of said SEQ ID NO. 5 or SEQ ID NO. 6 or SEQ ID NO. 7 or SEQ ID NO. 8 or the complement thereof and which comprises at least one CpG dinucleotide sequence; a fragment which hybridizes under moderately stringent or stringent conditions to at least 15 nucleotides of said SEQ ID NO. 9 or SEQ ID NO. 10 or SEQ ID NO. 11 or SEQ ID NO. 12 or the complement thereof and which comprises at least one CpG dinucleotide sequence; A fragment which hybridizes under moderately stringent or stringent conditions to at least 15 nucleotides of said SEQ ID NO. 13 or SEQ ID NO. 14 or SEQ ID NO. 15 or SEQ ID NO. 16 or the complement thereof and which comprises at least one CpG dinucleotide sequence.
  10. 10. An oligonucleotide for detecting colorectal cancer in vitro, comprising: SEQ ID NO. 17 and SEQ ID NO. 18.

Description

Composition for in vitro detection of colorectal cancer and application thereof Technical Field The application belongs to the field of molecular biology, relates to gene detection, and in particular relates to a composition for detecting colorectal cancer in vitro and application thereof. Background Colorectal cancer (CRC) is one of the most common malignant forms worldwide and severely threatens human health, with adenocarcinoma being the most common pathological form among colorectal cancers and also being the leading cause of death in patients. Surgical excision in combination with adjuvant chemotherapy is the primary treatment for early colorectal cancer patients to achieve long-term survival. Even if the patient receives surgical resection that appears to be successful, the risk of recurrence remains as photocopy. Although the operation can directly remove visible tumor tissues, it is difficult to ensure that all cancer cells are completely used up, and tiny residual focuses can be hidden everywhere in the body, so that the operation becomes a 'timing bomb' for recurrence in the future. Traditional imaging and serum tumor markers (e.g., CEA) have significant limitations in early recurrence monitoring, in that imaging can only detect macroscopic lesions, and CEA sensitivity and specificity are not ideal. Therefore, high-sensitivity molecular marker detection (such as ctDNA, methylation markers, etc.) after surgery has important clinical value. The novel biomarkers can detect tiny residual lesions (MRD) at a molecular level, early warn recurrence risk several months in advance than the traditional method, and strive for precious time window for clinical intervention. Meanwhile, the MRD detection can also realize accurate risk stratification, guide individualized auxiliary treatment decisions, avoid over treatment or insufficient treatment, and therefore remarkably improve survival prognosis of patients. Therefore, more accurate biomarkers and detection methods are urgently needed to better guide diagnosis and treatment of colorectal cancer and improve survival rate and quality of life of patients. Recent studies have shown that epigenetics play an important role in the development and progression of cancer. As an important mechanism of epigenetic science, DNA methylation regulation of various cancers has been intensively studied. Research data shows that the regulation of gene methylation is related to biological mechanisms such as chromatin structure, gene expression regulation and the like, that the change of cell gene methylation occurs in early stage of tumor formation and penetrates through the occurrence and development processes of cancer, and that the methylation of cancer suppressor genes is an important molecular mechanism for converting precancerous lesion tissues into malignant tumor cells. But currently there is a lack of detection techniques, methods and products for colorectal cancer methylation gene detection. Thus, there is a current need for methylation gene markers with high sensitivity and high specificity for colorectal cancer recurrence detection. Disclosure of Invention Based on the problems of the existing colorectal cancer detection, the application aims to provide a composition for detecting colorectal cancer in vitro, a kit and application thereof, and application for detecting colorectal cancer. The specific technical scheme of the application is as follows: 1. A composition for detecting colorectal cancer in vitro, the composition comprising: nucleic acid for detecting methylation status of a target gene, Wherein the methylation state of the target gene is characterized by methylation of a target sequence of the target gene, Wherein the target gene is selected from one or more than two of MSC gene, FOXE gene, GDF6 gene and C9orf50 gene. 2. The composition according to item 1, wherein the target sequence of the MSC gene is the sequence shown in any one of SEQ ID NOs 1 to 4 or comprises the sequence shown in any one of SEQ ID NOs 1 to 4, and/or The target sequence of FOXE gene is the sequence shown in any one of SEQ ID NOs 5-8 or contains the sequence shown in any one of SEQ ID NOs 5-8, and/or The target sequence of the GDF6 gene is or comprises the sequence shown in any one of SEQ ID NOs 9-12, and/or The target sequence of the C9orf50 gene is a sequence shown in any one of SEQ ID NOs 13-16 or comprises a sequence shown in any one of SEQ ID NOs 13-16. 3. The composition according to item 1 or 2, wherein the nucleic acid for detecting the methylation state of the target gene comprises: A primer which is a fragment of at least 9 nucleotides in a target sequence of the target gene, The fragment comprises at least one CpG dinucleotide sequence. 4. The composition of any one of items 1 to 3, wherein the nucleic acid for detecting methylation status of a target gene comprises: A probe which is a fragment of at least 15 nucleotides hybridised to the target sequence of the target gene