Prostate-specific antigen detection reagent and kit

A prostate-specific detection kit technology, applied in the field of immunoanalysis medicine, can solve the problems of high cost, complex reaction system, unsuitable for practical application, etc., and achieve the effect of high sensitivity, simple operation procedure and accurate detection

Active Publication Date: 2017-11-21
LINYI UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In the above method, in order to enhance the fluorescence detection signal, multiple DNA isothermal amplification cycle reactions are implemented, the reaction system used is complicated, the cost is high, and it is not suitable for practical application

Method used

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  • Prostate-specific antigen detection reagent and kit
  • Prostate-specific antigen detection reagent and kit
  • Prostate-specific antigen detection reagent and kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Example 1 [Configuration and testing and debugging of detection kit]

[0053] 1.1 Reagent configuration

[0054] PSA primary antibody, 20 times concentrated washing solution (0.02M phosphate buffer containing 0.05% Tween-20, pH=7.4), sample diluent (inactivated calf serum), containing fluorescein / quencher NEBuffer for labeled hairpin DNA strands.

[0055] 1.2 Detection and debugging steps

[0056] Prepare PSA series standard products with concentrations of 0, 0.2, 0.8, 1.6, 3.2, 8.6, 11.0, 20ng mL -1 .

[0057] (1) Coating of the microplate: Dissolve the PSA primary antibody with a coating buffer (50mM sodium carbonate, 50mM sodium bicarbonate, pH=9.6) to make the PSA primary antibody concentration 10μg / ml and 100μL / well Add to the microtiter plate and place overnight at 4°C. After discarding the coating solution the next day, wash with distilled water three times, add 150 μL of 1% bovine serum albumin to each well, and block at 37°C for 1 hour.

[0058] (2) Preparation of magn...

Embodiment 2

[0064] Example 2 [Using method of detection kit]

[0065] 1. Take out the kit from the refrigerator at 4°C, equilibrate at room temperature for 20 minutes, take a bottle of concentrated lotion, and add distilled water (1mL+19mL) for later use.

[0066] 2. Take the microplate out of the sealed bag, fill the wells with the washing liquid, and let it stand for 10-20 seconds, shake off the washing liquid, repeat the washing, and finally pat dry on a clean absorbent paper.

[0067] 3. Set a series of standard concentrations of 2 holes each, 2 holes for each sample hole, take 50μL each of the standard or sample solution and place in the corresponding plate hole, shake and mix well with a micro shaker, cover the reaction plate with a non-drying film, Then the reaction plate was incubated at 37°C for 30 minutes.

[0068] 4. Carefully shake off the reaction solution, and then wash five times as in step 2.

[0069] 5. Take 100 μL of the magnetic bead solution co-labeled with PSA secondary antibo...

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Abstract

The invention relates to a prostate-specific antigen detection reagent and a kit. The detection reagent comprises PSA first antigen, PSA second antigen and a magnetic bead commonly marked with shear enzymes, a signal amplification reaction liquid reacted with the shear enzymes. The signal amplification reaction liquid contains fluorescein/quenching agent marked block DNA chain; when the block DNA chain is cut off by the shear enzymes highly distributed on the magnetic bead, the fluorescein is separated from the quenching agent, thus the fluorescence signal can be released, thereby amplifying the detection signal of the PSA. The reagent and the kit are simple in operation, high in sensitivity, good in specificity, and applicable to rapid detection of PSA in seminal fluid and serum of a human body.

Description

Technical field [0001] The present invention relates to the field of immunoassay medicine, in particular, the present invention relates to a prostate specific antigen detection reagent and a kit containing the same. Background technique [0002] Prostatic Cancer (PCa) is currently a common cancer among men. If it can be diagnosed in an early stage, it can be effectively dealt with and treated. Prostate-specific antigen (PSA) has been the main protein biomarker for clinical diagnosis of prostate disease since it was developed decades ago. PSA is mainly present in serum as free PSA and bound PSA. The free state is called fPSA, and the bound state is called (tPSA) PSA. The level is 4ng mL -1 Or lower cases are considered normal values, and more than 4ng mL -1 It is considered a dangerous value. [0003] The traditional methods of detecting prostate specific antigen include enzyme-linked immunoassay (ELISA), radioimmunoassay (RIA), fluorescence method, chemiluminescence method, electr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/574
CPCG01N33/57434
Inventor 周宏刘静张宁波张书圣
Owner LINYI UNIVERSITY
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