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Immunofluorescence kit for detecting human prostate cancer antigens PSA and AR-V7 and application

A prostate cancer, immunofluorescence detection technology, applied in biological testing, fluorescence/phosphorescence, measurement devices, etc., can solve the problem that a single tissue sample is not enough to reflect tumor burden, cannot be dynamically monitored, and cannot reflect non-humoral rare and rare in real time Nuclear cell changes, etc.

Inactive Publication Date: 2018-12-11
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

[0010] 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 kit for detecting human prostate cancer antigens PSA and AR-V7 and application
  • Immunofluorescence kit for detecting human prostate cancer antigens PSA and AR-V7 and application
  • Immunofluorescence kit for detecting human prostate cancer antigens PSA and AR-V7 and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0078] Materials: smears of negatively enriched blood samples, 22RV1 and LNCaP smears for control cells.

[0079] Experimental steps:

[0080] 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;

[0081] 2. Wash slides with CYP1 for 3 min×3 times, 100-150 μL each time, to ensure that the entire sample area is covered;

[0082] 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 ice acetone:methanol (7:3) for 5 minutes, wash the slides with CYP1 for 3 minutes × 3 times , to absorb excess water;

[0083] 4. Add 100-150 μl of blocking solution to block at room temperature for 25-30 minutes. Absorb excess blocking solution, add 100 μl of diluted PSA antibody, AR-V7 antibody and CD45 antibody, and incubate in a humid chamber...

Embodiment 2

[0093] Materials: 1 tube of appropriate amount of anticoagulated blood, which is enriched by membrane filtration and then detected for protein. Experimental steps:

[0094] 1. Take an appropriate amount of peripheral blood and put it into a blood collection tube containing anticoagulant, and shake it slightly to mix.

[0095] 2. Add the suspension to the membrane filtration separation tumor cell technology device, and slowly pass through the filter and the filter membrane.

[0096] 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;

[0097] 4. Fix the cells on the filter membrane;

[0098] 5. Perform the same operation as in Example 1 to detect the protein.

Embodiment 3

[0100] Materials: 1 tube of appropriate amount of anticoagulated blood, which is enriched by microfluidic method and then detected for protein

[0101] Experimental steps:

[0102] 1. The appropriate amount of blood drawn is enriched using microfluidic chips of various principles.

[0103] 2. After enrichment, the samples were subjected to protein immunofluorescence detection.

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Abstract

The invention relates to an immunofluorescence kit for detecting human prostate cancer antigens PSA and AR-V7 and application. According to a method provided by the invention, a detection principle comprises the following steps: firstly, enriching non-body-fluid rare karyocytes; the detecting expression of target protein in enriched cells in the cells by adopting an immunofluorescence detection method according to an antigen and antibody reaction principle. The kit provided by the invention is used for carrying out fluorescence labeling on a common antigen CD45 of targeting cells and white blood cells, and positive and CD45 negative cells of the target protein are screened, so that the positive non-body-fluid rare karyocytes of specific protein in blood are judged and counted.

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 prostate cancer antigen PSA, androgen receptor splicing mutant AR-V7 immunofluorescence detection kit. The invention is suitable for detecting PSA and AR-V7 of different types of samples, and also relates to the use of the kit in detecting PSA and AR-V7. Background technique: [0002] Prostate cancer refers to epithelial malignant tumors that occur in the prostate gland. It is a common malignant tumor in men and is listed as one of the top five cancers that cause death in men worldwide. At present, androgen deprivation therapy is an important method for the treatment of prostate cancer, but most patients will develop castration-resistant prostate cancer (CRPC) within two years after treatment, and the prognosis is poor. [0003] Prostate specific antigen (PSA) is a serine protease. In 1987, PSA was approved by t...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/574G01N21/64
CPCG01N21/6486G01N33/57434G01N33/689
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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