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An aptamer-antibody method for detecting a trace of prostate-specific antigen (PSA) in blood and a kit

A specific technology for prostate cancer, applied in the field of protein detection, can solve the problems of high environmental purity requirements and high false positive rate of antibody specificity requirements

Inactive Publication Date: 2015-09-02
SUZHOU YOULIN BIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The first problem to be solved by the present invention is to overcome the problems of high requirements for environmental purity, very high requirements for antibody specificity and high false positive rate in the detection of specific antibodies for human prostate cancer-specific antigens in the prior art, and provide a blood An aptamer antibody detection method for prostate cancer-specific antigen, which is easy to use, sensitive, specific, reproducible, quick to produce results, does not require complex instruments and special skills, and will not appear between aptamers and target molecules (prostate cancer-specific Sexual antigens) and other non-specific proteins interfere with each other

Method used

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  • An aptamer-antibody method for detecting a trace of prostate-specific antigen (PSA) in blood and a kit
  • An aptamer-antibody method for detecting a trace of prostate-specific antigen (PSA) in blood and a kit
  • An aptamer-antibody method for detecting a trace of prostate-specific antigen (PSA) in blood and a kit

Examples

Experimental program
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Effect test

Embodiment 1

[0068] Example 1 Detection of prostate cancer cell protein samples and their controls

[0069] LNCaP (PSA+) cells were cultured in vitro (The Prostate2011, Vol71, Issue15, pg1668-79.), the adherent cells were digested with trypsin to suspend the cells, centrifuged to remove the culture medium, and physiological saline was added to prepare a single cell suspension. Count a certain amount of cells (10 7 )conduct experiment. According to the manufacturer's instructions (Abio), the total protein of LNCaP cells was extracted with the total cell protein extraction reagent, and the amount of quantitative total protein was measured. Then, the total protein was diluted with PBS buffer (pH7.2), and the equal volumes of solutions of each dilution contained 5 (Group A), 10 4 (Group B), 10 3 (Group C), 10 2 (Group D) and 10 1 (Panel E) Total protein of LNCaP prostate cancer cells.

[0070] According to the above method, the total protein of PC3 (PSA-) (The Prostate2011, Vol71, Issue...

Embodiment 2

[0071] Example 2: Comparison with the method of detecting prostate cancer-specific antigen by ELISA

[0072] According to the determined conditions, ELISA (Human PSA ELISA Kit, GenWay Biotech Inc) detection, according to the strength of the color reaction to judge the existence of the target protein molecule. Target molecule (PSA) expression negative PC3 cell total protein (A 0 , B 0 、C 0 、D 0 ,E 0 Group diluent) is the corresponding control group. The results are as follows Figure 1 It shows that the OD value of the total protein of PC3 (PSA-) cells with negative target molecule expression is very low, which can be used as a background control, and its detection line is used as the standard baseline of detection (higher than this line is a positive test result); In the triplicate detection of LNCaP cell serial dilution protein samples known to contain the target molecule PSA protein, even the lower dilution D sample (10 2 cells / ml), and the OD value was also significa...

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Abstract

An aptamer-antibody method for detecting a trace of prostate-specific antigen (PSA) in blood is provided. A specific PSA aptamer coupled to magnetic particles and a specific PSA antibody coupled to nanogold particles are respectively mixed with a sample to be detected so as to detect, identify and / or quantify a target molecule (the PSA). The method is simple and convenient to use, sensitive, high in specificity, good in repeatability, rapid in result production, and free of needs for complex instruments and special techniques, and can be popularized to actual industrial application.

Description

technical field [0001] The invention relates to the technical field of protein detection methods, in particular, the invention relates to a method for detecting, identifying and / or quantifying trace prostate cancer-specific antigens in human blood. In addition, the present invention also relates to kits used in the method and applications thereof. Background technique [0002] Prostate cancer is one of the most common fatal cancers in elderly men. Surgical resection is the first choice in the comprehensive treatment of prostate cancer, but the recurrence rate and metastasis rate after resection of prostate cancer are high: cancer cells are easy to enter the peripheral blood from the primary tumor Tumor metastasis occurs, which is the main cause of treatment failure in prostate cancer. A large number of experiments have confirmed that the detection of Circulating Tumor Cells (CTCs) in circulating blood will be helpful for the early diagnosis of prostate cancer, the monitorin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/574G01N33/553
CPCG01N33/57434
Inventor 李为李宏
Owner SUZHOU YOULIN BIO TECH
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