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Time-resolved fluorescence immunoassay kit for detecting t-PAI-C and application

A kit, t-pai-c technology, applied in the field of immunoassay analysis and nanobiology, can solve the problems of unreliability and achieve the effects of improving efficiency, shortening detection time, and long decay time

Pending Publication Date: 2020-11-17
ZHEJIANG SHENGYU MEDICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] At present, there is no reliable commercial t-PAI-C detection method, and ELISA assay kits are mostly used in experiments. The detection principle is: the kit uses a double-antibody one-step sandwich enzyme-linked immunosorbent assay (ELISA)

Method used

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  • Time-resolved fluorescence immunoassay kit for detecting t-PAI-C and application
  • Time-resolved fluorescence immunoassay kit for detecting t-PAI-C and application
  • Time-resolved fluorescence immunoassay kit for detecting t-PAI-C and application

Examples

Experimental program
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Effect test

Embodiment 1

[0051] The preparation of embodiment 1 kit

[0052] A time-resolved fluorescent immunoassay kit for detecting tissue plasminogen activator / plasminogen activator inhibitor-1 complex (t-PAI-C), the kit includes the following components:

[0053] The preparation of described t-PAI-C calibrator solution: get the t-PAI-C solution of high concentration with the NaN containing the BSA of 2g / L and 1g / L 3 The 50mmol / L Tris-HCl reaction buffer solution with a pH of 7.8 was used to prepare calibrator solutions with different concentrations, which were subpackaged, freeze-dried, and stored at 2-8°C.

[0054] The preparation of the magnetic particle solution coated with t-PAI-C antigen: take ferric oxide microspheres with a diameter of 50-5000nm and activate them with glutaraldehyde with a mass concentration of 5-10%, and mix them at room temperature for 4 hours Finally, wash 1-3 times with 0.05mol / L PBS buffer solution with a pH of 7.2, and then suspend with the above buffer solution to ...

Embodiment 2

[0060] A time-resolved fluorescent immunoassay kit for detecting t-PAI-C, such as figure 1 , 2 As shown, it consists of a box body 1 , a cover body 2 covering the top opening of the box body 1 , and a battery pack 11 located below the bottom of the box body 1 .

[0061] Such as Figure 4 As shown, the inside left side of the box body 1 is provided with a storage block 3, and the upper surface of the storage block 3 has two rows of storage holes 5 for storing freeze-dried cups 4, and the right side of the box body 1 is provided with a storage slot 6, and the storage slot 6 The opposite two sides are provided with support strips 23, and the upper end surfaces of the support strips 23 are provided with lifting plates 24, and the upper end surfaces of the lifting plates 24 are placed with reagent racks 8 for storing reagent strips 7, such as Figure 7 As shown, the reagent strip 7 includes a reagent plate 9 that is detachably connected to the reagent rack 8. The upper end of the...

Embodiment 3

[0069] A method for detecting tissue plasminogen activator / plasminogen activator inhibitor-1 complex (t-PAI-C) using the time-resolved fluorescent immunoassay kit described in Example 1, specific steps as follows:

[0070]1. After the calibrator solution is calibrated, dilute the serum to be tested with the reaction buffer 1:100 and take 100ul, mix it with the 50ul magnetic particle solution coated with the t-PAI-C antibody, react for 20 minutes, and perform magnetic separation , washed twice with the cleaning solution, adding the europium-labeled goat anti-human IgG antibody solution diluted with the reaction buffer solution into the mixed magnetic particle solution, incubating for 20 minutes, and then washing with the cleaning solution for magnetic separation , repeating the above cleaning step 4 times, and then adding the enhancing solution;

[0071] 2. The fully automatic time-resolved fluorescence immunoassay analyzer respectively measures the fluorescence value of europ...

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Abstract

The invention relates to a biological detection kit, in particular to a time-resolved fluorescence immunoassay kit for detecting a tissue plasminogen activator / plasminogen activator inhibitor 1 compound (t-PAI-C) and application, and belongs to the technical field of immunoassay analysis and nano-biology. The time-resolved fluoroimmunoassay kit for detecting a tissue plasminogen activator / plasminogen activator inhibitor 1 compound (t-PAI-C) comprises a reaction buffer solution, a cleaning solution, an enhancement solution, a t-PAI-C antigen-coated magnetic particle solution, a europium-labeledgoat anti-human IgG antibody solution and a t-PAI-C freeze-dried calibration product which are integrated on a reagent strip. The kit provided by the invention can provide a reaction system close toa homogeneous phase, detect the content of tPAIC, shorten the detection time, improve the efficiency and provide convenience for clinical use.

Description

technical field [0001] The invention relates to a biological detection kit, in particular to a time-resolved fluorescence immunoassay kit for detecting tissue plasminogen activator / plasminogen activator inhibitor-1 complex (t-PAI-C) and applications, belonging to the field of immunoassay analysis and nanobiotechnology. Background technique [0002] The key link in venous thrombosis is the generation of thrombin, which acts on fibrinogen to convert fibrin to form thrombus. Since the generated thrombin has a very short half-life in the blood, only a few seconds, and is quickly neutralized by anticoagulant substances, it is very difficult to directly measure thrombin. Instead, the detection of thrombin and its inhibitor antithrombin ( AT) 1:1 bound thrombin-antithrombin complex (TAT). The production of TAT directly confirms the activation of the coagulation system. Thrombomodulin (TM) is expressed on the surface of vascular endothelial cells and can assist the thrombin-activ...

Claims

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Application Information

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IPC IPC(8): G01N33/573G01N33/68G01N33/58G01N33/543G01N33/533
CPCG01N33/573G01N33/6893G01N33/582G01N33/54326G01N33/54346G01N33/533G01N2333/96433G01N2333/8132G01N2800/226G01N2800/324
Inventor 赵蕾石坚盛誉盛炯
Owner ZHEJIANG SHENGYU MEDICAL TECH CO LTD
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