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Colorectal cancer early screening kit

A colorectal cancer and kit technology, applied in the field of molecular biology, can solve the problems of affecting the effect of treatment and prognosis, easy contamination, high false positive rate, avoid false negative and false positive results, improve specificity and accuracy Sexuality, the effect of improving the cure rate and survival rate

Inactive Publication Date: 2017-12-15
SUREXAM BIO TECH
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  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the radical treatment of colorectal cancer is still the first surgical treatment. The five-year survival rate of patients with colorectal cancer detected in the early stage is as high as 90%. Once metastasized, the survival rate of colorectal cancer patients is less than 10%.
However, the current early diagnosis rate of colorectal cancer is less than 40%, and according to relevant statistics, the misdiagnosis rate of colorectal cancer is 30%, which seriously affects the effect of treatment and prognosis
Although there are also early screening technologies for colorectal cancer-related tumor gene levels, such as marker gene methylation site detection, microRNA detection, etc., but the current detection is mainly based on the principle of PCR reaction, which has defects such as easy contamination and high false positive rate.
In addition, the lack of specific molecular markers for early diagnosis is also an important factor limiting the early diagnosis of colorectal cancer

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] There are two types of colorectal cancer early screening kits described in this example, with labeled probes and without labeled probes.

[0027] Among them, the colorectal cancer early screening kit A with labeled probes mainly includes:

[0028] 1. Capture probe

[0029] The capture probe consists of three parts, the 5' end to the 3' end are sequentially the P1 sequence that can be complementary to the mRNA of the corresponding target gene, the spacer sequence, and the P2 sequence that is complementary to the corresponding amplification probe P3 sequence, Target genes of the same category have the same P2 sequence in their capture probes. The spacer is used to space the capture probe P2 sequence from the target mRNA, and by setting a spacer sequence of appropriate length inside the probe, it can reduce steric hindrance, improve the efficiency of the hybridization reaction and the specificity of the hybridization reaction . The spacer arm of the capture probe of the...

Embodiment 2

[0050] Example 2 Using kit A in Example 1 to detect circulating tumor cells in peripheral blood of patients with colorectal cancer

[0051] The formula of described various solutions is as follows:

[0052]

[0053]

[0054] The probe mixture, amplification mixture, and chromogenic mixture in this example all use all the probes in the corresponding gene list of Colorectal Cancer Early Screening Kit A with labeled probes in Example 1.

[0055] 1. Sample pretreatment, filter the screening cells onto the filter membrane

[0056] 1. Preserve the blood sample in the sample preservation tube with preservation solution, centrifuge at 600×g for 5 minutes, discard the supernatant, and remove the red blood cells.

[0057] 2. Add 4mL PBS and 1mL fixative, vortex to mix, and let stand at room temperature for 8min.

[0058] 3. Sample filtration: Transfer the liquid in the sample storage tube to the filter, turn on the vacuum filtration pump to pump out the liquid; add 4mL PBS to th...

Embodiment 3

[0108] The selection of embodiment 3 target detection gene quantity and kind

[0109] 1. Design of kit preparation (selection of the number and types of target detection genes)

[0110] The colorectal cancer screening genes of the kit of the present invention are selected from: CK20, ZEB1, CEA, and CD24, and the corresponding selection is made according to the experimental group. The genes related to non-blood cells: EGFR, CDX2, MACC1, and LGR5; the exclusion genes: CD146 , CD105, VWF, CD45 and CD34.

[0111] For the selection of colorectal cancer screening genes, refer to experimental groups 1-4, select one, two, three and four target genes respectively, and compare their detection effects, and use all of the non-blood-derived cell-related genes and exclusion genes The specific design of the target gene is shown in Table 7.

[0112] The compositions and quantities of the capture probes, amplification probes and labeling probes, detection methods, etc. of each group of corre...

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Abstract

The invention relates to a colorectal cancer early screening kit. The kit comprises capture probes, amplification probes and labeled probes for detection of target gene mRNAs. The target genes comprise colorectal cancer screening genes and non-blood cell related genes. The colorectal cancer screening genes comprise at least two of CK20, ZEB1, CEA and CD24. The non-blood cell related genes comprise at least two of EGFR, CDX2, MACC1 and LGR5. Through comprehensive assessment and statistical analysis and screening in lot of experiments, the colorectal cancer screening genes, the non-blood cell related genes and exclusion genes are selected so that detection of a single target gene is realized and simultaneous detection of multiple target genes is realized and thus the screening cells can be completely detected, leukocytes, hematopoietic stem cells and endothelial cells in blood are distinguished from the screening cells, false negative and false positive results are avoided and the accuracy of detection is further improved.

Description

technical field [0001] The invention belongs to the field of molecular biology, relates to medicine and biotechnology, in particular to a colorectal cancer early screening kit. Background technique [0002] Colorectal cancer is a cancer formed by abnormal growth of cells in the colon or rectum. It is a common malignant tumor in the gastrointestinal tract. Symptoms are not obvious, and generally it will go through the evolution process from normal mucosal tissue, adenoma, to malignant tumor in 7 to 10 years. In recent years, the global incidence of colorectal cancer has risen sharply. In recent years, more than one million people die of colorectal cancer every year. Colorectal cancer mostly occurs in men over middle age, with a male to female incidence ratio of about 2:1, and most patients are over 40 years old. Early colorectal cancer lacks specific symptoms, and when patients seek treatment due to obvious clinical symptoms, they are often in the middle and advanced stages...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6841C12Q1/6886C12Q2600/158C12Q2563/107C12Q2565/543
Inventor 刘苏燕吴诗扬董艳
Owner SUREXAM BIO TECH
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