Joint detection kit for multiple markers on CTCs (circulating tumor cells) and application of kit

A technology for combined detection of tumor cells, applied in the determination/testing of microorganisms, biochemical equipment and methods, etc., can solve the problems of high detection cost, unsuitable for high-throughput detection, etc., to avoid interference and not require high equipment , the effect of simple operation

Inactive Publication Date: 2018-01-16
ZHONGSHAN HOSPITAL FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

As a new chip technology, CTC-chip has the advantages of high sensitivity and good specificity, but it requires

Method used

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  • Joint detection kit for multiple markers on CTCs (circulating tumor cells) and application of kit
  • Joint detection kit for multiple markers on CTCs (circulating tumor cells) and application of kit
  • Joint detection kit for multiple markers on CTCs (circulating tumor cells) and application of kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] A combined detection kit for multiple stem cell markers on the surface of circulating tumor cells, consisting of the following reagents:

[0062] 1. Peripheral blood circulating tumor cell enrichment reagent: RosetteSep Human CD45Depletion Cocktail (Stemcell, Canada), 5*2ml, Cat.No.15162;

[0063] 2. Mini Kit (50) (Qiagen, Germany), Cat. No. 74104;

[0064] 3. Reverse Transcription Kit (Qiagen, Germany), Cat. No. 205313;

[0065] 4. SuperMix-UDG (Invitrogen, USA), Cat. No. C11730-025;

[0066] 5. Taq-Man probe (self-made, 10mM):

[0067] EpCAM probe: FAM-ACGGCGACTTTTGCCGCAGCT-TAM;

[0068] CD133probe:FAM-ACCAGCGTGAAAACTAGCCTGCGGTC-TAM;

[0069] CD90probe:FAM-TCAGCCTGACCCGTGAGACAAAGAAGC-TAM;

[0070] CK19probe: FAM-CCGCTCCGGGCACCGATCT-TAM;

[0071] β-actin probe: FAM-ATCACTGCCCTGGCACCCAGCA-TAM;

[0072] 6. Specific primers (self-made, 10mM):

[0073] EpCAM-F: 5'-CTCGCGTTCGGGCTTCT-3';

[0074] EpCAM-R: 5'-TGTAGTTTTCACAGACACATTCTTCCT-3';

[0075] CD133-F: 5...

Embodiment 2

[0085] A combined detection kit for multiple stem cell markers on the surface of circulating tumor cells, the composition of which is the same as in Example 1.

[0086] Use the above-mentioned circulating tumor cell detection kit to detect circulating tumor cells, and evaluate the CTC recovery rate of the established method. For specific steps, please refer to the independent patent (patent number: 2014100815089.9).

Embodiment 3

[0088] A combined detection kit for multiple stem cell markers on the surface of circulating tumor cells, the composition of which is the same as in Example 1.

[0089] Use the above-mentioned circulating tumor cell detection kit to detect circulating tumor cells, and evaluate the imprecision within batches and batches. For specific steps, please refer to the independent patent (patent number: 2014100815089.9).

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PUM

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Abstract

The invention provides a joint detection kit for multiple stem cell markers on surfaces of CTCs (circulating tumor cells) and an application of the kit. The joint detection kit for multiple stem cellmarkers on surfaces of CTCs comprises 1) a peripheral blood CTC enrichment reagent; 2) an RNA extraction reagent; 3) an RNA reverse transcription reagent; 4) a fluorescence labeled Taqman probe and aspecific primer; 5) a qPCR reaction system reagent and a hepatocellular carcinoma prediction formula. The kit can be used for biopsy of minimally invasive fluids of tumor patients, early diagnosis andearly warning of tumor metastasis and relapse are realized, the anti-tumor therapy effect and the tumor progression condition are monitored, molecular subtyping based on CTCs and screening of specific sensitive drugs for the CTCs are realized, so that knowledge about tumor metastasis, relapse and drug resistance related mechanisms is improved, individualized medical treatment of tumor patients isguided, the overall survival of the tumor patients is significantly improved, and the kit has great clinical significance.

Description

technical field [0001] The invention relates to a kit for combined detection of multiple stem cell markers on the surface of circulating tumor cells and an application thereof. Background technique [0002] In recent years, the morbidity and mortality of hepatocellular carcinoma have risen rapidly, ranking fifth among malignant tumors. Although treatment techniques have been greatly improved, the 5-year survival rate has not improved significantly, and about 50% of patients have tumor recurrence or distant metastasis 5 years after surgery. Therefore, predicting the postoperative recurrence of liver cancer patients for early necessary intervention is the key to further improve the prognosis of liver cancer. There is an urgent need for an effective and reliable detection technology to solve the problems faced in the treatment of HCC such as early diagnosis of disease, screening of high-risk recurrence, evaluation of anti-tumor treatment effect, prevention and control of dista...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886C12Q1/686C12Q1/02
Inventor 樊嘉杨欣荣孙云帆郭玮周俭
Owner ZHONGSHAN HOSPITAL FUDAN UNIV
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