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Quantitative determination kit for human epididymis secretory protein 4

A technology for quantitative determination of epididymal secreted proteins, applied in biological tests, measuring devices, analytical materials, etc., can solve the problem of less than 25% of early diagnosis, and achieve the effects of no radioactive pollution, strong specificity and high sensitivity

Inactive Publication Date: 2014-12-24
GUANGZHOU DARUI BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

About 90% of stage I ovarian cancer can survive for a long time after treatment, but less than 25% of stage I ovarian cancer can be diagnosed early

Method used

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  • Quantitative determination kit for human epididymis secretory protein 4
  • Quantitative determination kit for human epididymis secretory protein 4
  • Quantitative determination kit for human epididymis secretory protein 4

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1 Composition of the kit

[0047] 1) M reagent: 1 bottle (100 test pack: 7.0 mL) 6F8 antibody-coated magnetic microspheres, stored in 0.1M Tris buffer containing 2% BSA stabilizer. Preservative: 0.05% NaN 3 .

[0048] 2) R1 reagent: 1 bottle (100 test pack: 6.0 mL) analysis buffer, stored in 0.1M Tris buffer containing 2% BSA stabilizer. Preservative: 0.05% NaN 3 .

[0049] 3) R2 reagent: 1 bottle (100 test pack: 7.0 mL) of H4 antibody labeled with acridine salt, stored in 0.5% Tris buffer containing 0.2% BSA stabilizer. Preservative: 0.01% NaN 3 .

[0050] 4) Pre-excitation solution: 1.32% (w / v) hydrogen peroxide.

[0051] 5) Excitation solution: 0.35N sodium hydroxide.

[0052] 6) Washing solution: 0.05% Tween-20 0.1M phosphate buffer solution.

[0053] 7) HE4 calibrator: Calibrator 1: 10pmolL -1 ; Calibrator 2: 300pmolL -1 . That is, the two concentrations of HE4 antigen were stored in 0.1M Tris buffer containing 2% BSA stabilizer. Preservative: 0.05% NaN 3 .

[005...

Embodiment 2

[0056] Example 2 Method of using the kit

[0057] 1. Load reagents

[0058] 1.1M reagent is yellow-brown magnetic microsphere suspension, precipitation is normal. Before loading the HE4 quantitative determination reagent on the Caris200 system for the first time, the M reagent bottle needs to be turned and mixed.

[0059] 1.2 If the magnetic microspheres are still attached to the bottle wall by visual inspection, it is necessary to continue turning over until the magnetic microspheres are resuspended. If there is no suspension for a long time, the reagent cannot be used.

[0060] 1.3 Load the HE4 reagent on the Caris200 according to the requirements of the instrument to ensure that all the reagents required for the test are on the machine. Ensure that all reagent bottle caps have been removed.

[0061] 2. Calculate the required sample size

[0062] 2.1 When performing HE4 detection for the first time, 175μL of sample needs to be added, and 25μL of sample needs to be added to the same...

Embodiment 3

[0079] Example 3 Establishment and optimization of reaction system

[0080] Optimization of the reaction mode: Use 6F8 as the coating antibody and H5 as the detection antibody to measure the standard, using the one-step method and the two-step method respectively. The difference between the two is that the one-step method combines the coating antibody standard and the detection antibody at the same time Add to the reaction well, incubate at 37°C for 25min and wash 4 times. Add the pre-excitation solution and test the luminous intensity while adding the excitation solution. In the two-step rule, add the coated antibody and standard product and incubate at 37°C for 15min, wash 2 After the second time, add the detection antibody, incubate for 10 minutes, wash twice, add the pre-excitation solution and add the excitation solution while testing on the machine to measure the luminescence intensity. Use the logarithm of the antigen concentration as the abscissa and the logarithm of the...

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Abstract

The invention provides a quantitative determination kit for human epididymis secretory protein 4 (HE4), which is applicable to a full-automatic chemiluminescence immunoassay analyzer, in particular relates to a preparation process of a kit for quantitatively detecting the concentration level of HE4 in human serum by adopting a dual-antibody sandwich acridinium ester-marked tubular chemiluminescence method. The kit adopts the dual-antibody sandwich immunoassay and utilizes a chemiluminescence magnetic microsphere immune technology, magnetic microspheres which are coated with anti-HE4 antibodies are specifically combined with the HE4 in a standard product / sample in a reaction cup and then react with another anti-HE4 antibody marked by the acridinium ester to form an immune complex, and the immune complex has the acid-alkali chemical reaction in a pre-trigger solution and a trigger solution, so that relative light unit (RLU / s) of the chemiluminescence reaction can be measured; the content of the HE4 in the standard product / sample is in direct proportion to the relative light unit (RLU / s) measured by an optical system, and the content of HE4 in a serum sample can be quantitatively measured by virtue of standard curve fitting. The quantitative determination kit has advantages of high sensitivity, high specificity, good stability, simplicity and convenience in operation, low cost and the like.

Description

Technical field [0001] The invention relates to a kit for the quantitative determination of human epididymis secretory protein 4 (HE4) suitable for an automatic chemiluminescence immunoassay analyzer Caris 200, in particular to a double antibody sandwich acridinium ester labeled tubular chemiluminescence method The preparation process of the kit for quantitatively detecting the concentration of HE4 in human serum. Background technique [0002] The incidence of ovarian cancer in gynecological tumors is second only to cervical cancer and uterine body cancer, while its fatality rate is the first. This is mainly because the ovaries are hidden deep in the bone cavity, and the early symptoms of ovarian cancer are not obvious, and these symptoms are easily confused with other non-cancerous conditions. More than 70% of patients are already in advanced stages when they see a doctor. Therefore, ovarian cancer has been Called the "silent killer". The current diagnostic methods for ovarian...

Claims

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Application Information

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IPC IPC(8): G01N33/68
CPCG01N33/57488G01N33/54326G01N33/57449G01N33/6893G01N2800/50G01N2800/52
Inventor 徐伟文张琼余娟平李志雄吴英松李明
Owner GUANGZHOU DARUI BIOTECH
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