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Colorectal cancer diagnosis kit

A colorectal cancer, kit technology, applied in biochemical equipment and methods, DNA/RNA fragment, microorganism determination/inspection, etc. The effect of reducing error rates and overcoming limitations

Inactive Publication Date: 2019-10-29
GUANGZHOU ZHONGYU BIOTECHNOLOGY CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are few studies on miRNA methylation in colorectal cancer
The role of miR-17-92 cluster methylation level in tumors has not been reported

Method used

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  • Colorectal cancer diagnosis kit
  • Colorectal cancer diagnosis kit
  • Colorectal cancer diagnosis kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1 Screening of methylation sites (CpG sites)

[0037] The miR-17-92 cluster was analyzed using the University of California Santa Cruz (UCSC) database, and bisulfite sequencing (BSP) primers were designed to find its CpG sites, namely methylation sites. The miR-17-92 cluster CpG sites were concentrated between the upstream 3.5kb sequence and the downstream 2kb sequence of the miR-17-92 cluster transcription initiation site. figure 1 The CpG site between the upstream 2kb sequence of the transcription start site and the downstream 2kb sequence is shown, and the present invention selects the miR-17-92 cluster transcription start site upstream 1.5kb sequence to the downstream 1.5kb sequence (SED ID NO:25 )conduct experiment.

Embodiment 2

[0038] Example 2 Primer Design

[0039] The sequencing primers were designed as bisulfite sequencing (BSP) primers. The primer sequences are listed in Table 1.

[0040] Table 1 Primer sequences for methylation sites

[0041]

Embodiment 3

[0042] Embodiment 3 sample collection

[0043] The feces, colorectal cancer tissues and paracancerous tissues of 204 colorectal cancer patients were collected. The surgical resection specimens of all cases were subjected to routine pathological examination to confirm the diagnosis of cancer. The DNA or RNA was extracted in the laboratory, and the cDNA and DNA after RNA reverse transcription were stored at -20°C; and 164 age-matched normal controls were selected as controls for stool testing during the same period. All patients were treated surgically and did not receive radiotherapy or chemotherapy before operation.

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Abstract

The invention belongs to the field of biotechnology, and particularly relates to a colorectal cancer diagnosis kit. The kit contains a reagent for detecting the methylation level of a to-be-detected sequence in a to-be-detected sample, and the to-be-detected sequence is located between an upstream 3.5kb sequence and a downstream 2kb sequence of an miR-17-92 cluster transcriptional start site. It is found for the first time that the low methylation level of an original cancer non-coding RNA miR-17-02 cluster in colorectal cancer is relevant to the generation and development of the colorectal cancer. The methylation level of the miR-17-92 cluster can be used as a biological indicator for tumor detection to assist diagnosis of the colorectal cancer, so that detection specificity and sensitivity are improved, the error rate is lowered, and the quite important clinic guidance significance to the tumor pathogenesis and diagnosis is achieved.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a diagnostic kit for colorectal cancer. Background technique [0002] Colorectal cancer (CRC) is one of the most common cancers in the world. The incidence of colorectal cancer in my country is about 14.2 / 100,000, showing a sharp upward trend in the past 20 years, and has become one of the fastest-rising malignant tumors. The occurrence of colorectal cancer must go through the normal mucosa, hyperplasia, adenoma formation, adenoma canceration, and the staged evolution of invasion and metastasis, and its development process can last for more than 10 years. Among them, adenomatous polyps are the most important precancerous lesions of colorectal cancer. Adenomatous polyps gradually increase in size and often take years to eventually become malignant. If colorectal cancer can be detected in the early stage, after surgical resection, the cure rate is very high, which can reach more tha...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12N15/11
CPCC12Q1/6886C12Q2600/154C12Q2600/178
Inventor 王淋立何敏仪莫健王萍陈帝武
Owner GUANGZHOU ZHONGYU BIOTECHNOLOGY CO LTD
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