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Immunofluorescence kit for detecting ER and PR antigen and application

A technology for immunofluorescence detection and antigen, which is applied in biological testing, material inspection products, etc., and can solve problems such as the inability to identify the expression state

Inactive Publication Date: 2018-11-09
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

In this study, reverse transcription PCR was used to detect ER and PR. RNA is easily degraded, and the expression status of ER and PR in single cells cannot be identified.

Method used

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  • Immunofluorescence kit for detecting ER and PR antigen and application
  • Immunofluorescence kit for detecting ER and PR antigen and application
  • Immunofluorescence kit for detecting ER and PR antigen and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0082] Materials: smears of negatively enriched blood samples, and RT-474 cell smears for control cells. Experimental steps:

[0083] 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;

[0084] 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;

[0085] 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 moisture;

[0086] 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 ER antibody, PR antibody and CD45 antibody, and incubate in a humid chambe...

Embodiment 2

[0095] Materials: 1 tube of appropriate amount of anticoagulated blood, which is enriched by membrane filtration and then detected for protein. Experimental steps:

[0096] 1. Take an appropriate amount of peripheral blood and put it into a blood collection tube containing anticoagulant, and shake it slightly to mix.

[0097] 2. Add the suspension to the membrane filtration separation tumor cell technology device, and slowly pass through the filter and the filter membrane.

[0098] 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;

[0099] 4. Fix the cells on the filter membrane;

[0100] 5. Perform the same operation as in Example 1 to detect the protein.

Embodiment 3

[0102] Materials: 1 tube of appropriate amount of anticoagulated blood, which is enriched by microfluidic method and then detected for protein. Experimental steps:

[0103] 1. The appropriate amount of blood drawn is enriched using microfluidic chips of various principles.

[0104] 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 ER and PR antigen and application. The detection principle of the method disclosed by the invention is as follows: firstly enriching non-body fluid rare karyocyte, and then detecting expression of interest protein in the enriched cell in the cell by adopting an immunofluorescence detection method according to an antigen antibody reaction principle. The kit can perform identification and counting on the positive non-body fluid rare karyocyte of specific protein in blood by screening the positive and CD45 negative cells of the interest protein.

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 ER and PR antigen immunofluorescence detection kit, which is suitable for detection of ER and PR antigens such as breast cancer and endometrial cancer, and also relates to Use of the kit in detecting ER and PR antigens. Background technique: [0002] At present, the incidence of cancer is relatively high, which seriously threatens human health. Some cancers are difficult to detect early and are often diagnosed at an advanced stage. Correspondingly, the 5-year survival rate of advanced cancer is significantly lower than that of early-stage cancer, and early detection, early diagnosis and precise treatment can significantly prolong the survival time of patients. [0003] Breast cancer and endometrial cancer often have high expression of estrogen receptor (Estrogen receptor, ER) and progesterone receptor (Progester...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68
CPCG01N33/68
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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