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Immunofluorescence detection kit for androgen receptor splicing variant AR-V7 and application

An immunofluorescence detection, AR-V7 technology, applied in the field of medical detection, can solve the problems of not being able to effectively reflect the changes of non-humoral rare nucleated cells in real time, repeated sampling is difficult, and a single tissue sample is not enough to reflect the tumor burden.

Inactive Publication Date: 2019-01-11
北京莱尔生物医药科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] At present, the pathological detection of tumor patients is mainly histological detection, which is faced with the difficulty of repeated sampling, cannot be dynamically monitored, and a single tissue sample is not enough to reflect the overall tumor burden, and cannot effectively reflect the non-humoral rare nuclei entering the body fluid in real time. cell changes

Method used

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  • Immunofluorescence detection kit for androgen receptor splicing variant AR-V7 and application
  • Immunofluorescence detection kit for androgen receptor splicing variant AR-V7 and application
  • Immunofluorescence detection kit for androgen receptor splicing variant AR-V7 and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0076] Materials: smears of negatively enriched blood samples and 22RV1 smears for control cells.

[0077] Experimental steps:

[0078] 1. Draw 3.5ml of peripheral blood into an ACD (sodium citrate) anticoagulant tube. use The human peripheral blood leukocyte depletion kit negatively enriches tumor cells and fixes them on glass slides;

[0079] 2. Wash slides with CYP1 for 3 minutes x 3 times, 100-150 μL each time, to ensure that the entire sample area is covered;

[0080] 3. Absorb the excess liquid on the slide, add CYPP for 5 minutes, wash the slides with CYP1 as above for 3 minutes × 1 time; absorb excess liquid, add 200 μl of ice acetone:methanol (7:3) for 5 minutes, and wash the slides with CYP1 for 3 minutes × 3 times , to absorb excess water;

[0081]4. Add 100-150 μl of blocking solution to block at room temperature for 25-30 minutes. Suck off excess blocking solution, add 100 μl of diluted AR-V7 antibody and CD45 antibody, and incubate in a humid chamber at roo...

Embodiment 2

[0087] Materials: 1 tube of appropriate amount of anticoagulated blood, which is enriched by membrane filtration and then detected for protein. Experimental steps:

[0088] 1. Take an appropriate amount of peripheral blood and put it into a blood collection tube containing anticoagulant, and shake it slightly to mix.

[0089] 2. Add the suspension to the membrane filtration separation tumor cell technology device, and slowly pass through the filter and the filter membrane.

[0090] 3. After the filtration is completed, continue to add 50ml of 0.01M PBS to the membrane filtration device, wash the cell suspension attached around the tube wall into the membrane filtration device, and let it pass through the filter and membrane;

[0091] 4. Fix the cells on the filter membrane;

[0092] 5. Perform the same operation as in Example 1 to detect the protein.

Embodiment 3

[0094] Materials: 1 tube of appropriate amount of anticoagulated blood, which is enriched by microfluidic method and then detected for protein. Experimental steps:

[0095] 1. The appropriate amount of blood drawn is enriched using microfluidic chips of various principles.

[0096] 2. After enrichment, the samples were subjected to protein immunofluorescence detection.

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Abstract

The invention relates to an immunofluorescence detection kit for an androgen receptor splicing variant AR-V7 and an application. The detection principle of the method according to the invention is asfollows: firstly, enriching non-humoral rare nucleated cells, and then adopting an immunofluorescence detection method for detecting expression of target proteins in the enriched cells according to anantigen antibody reaction principle. The kit judges and counts positive non-humoral rare nucleated cells of specific protein in blood by performing fluorescence labeling on target cell and leukocytecommon antigen CD45 and screening target protein positive and CD45 negative cells.

Description

Technical field: [0001] The invention relates to a medical detection method, in particular to the detection of tumor cells in body fluids, in particular to a human androgen receptor splicing mutant AR-V7 immunofluorescence detection kit. The invention is suitable for detecting the androgen receptor splicing mutant AR-V7 in different types of samples, and also relates to the use of the kit in detecting the androgen receptor splicing mutant AR-V7. Background technique: [0002] Prostate cancer refers to epithelial malignant tumors that occur in the prostate gland, and is listed as one of the top five cancers that cause death in men worldwide. At present, an important method for the treatment of prostate cancer is androgen deprivation therapy, which uses certain specific inhibitors to block the androgen receptor and its signaling pathways, thereby prolonging the survival of patients. However, most patients will develop castration-resistant prostate cancer (CRPC) within two yea...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/574G01N33/68G01N33/533G01N21/64
CPCG01N21/6428G01N33/533G01N33/57434G01N33/57496G01N33/6803G01N2800/7028
Inventor 郭素杰郭志敏樊晓婷
Owner 北京莱尔生物医药科技有限公司
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