Kit for joint detection of ovarian cancer tumor markers HE4 and CA125 as well as preparation method and application thereof

A technology of CA125 and tumor markers, applied in the field of immunoassay, can solve the problems of short reaction time and less reagent consumption

Active Publication Date: 2018-05-08
北京惠中医疗器械有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] So far, there is a lack of a magnetic particle immunoassay in the clinical detection of ovarian cancer. The detection process is simple, the reaction time is short, the reagent consumption is small, and the detection process is ...

Method used

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  • Kit for joint detection of ovarian cancer tumor markers HE4 and CA125 as well as preparation method and application thereof
  • Kit for joint detection of ovarian cancer tumor markers HE4 and CA125 as well as preparation method and application thereof
  • Kit for joint detection of ovarian cancer tumor markers HE4 and CA125 as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] This example provides a kit for joint detection of ovarian cancer tumor markers HE4 and CA125, including calibrator, magnetic particles jointly labeled with HE4 antibody and CA125 antibody, enzyme label, chemiluminescent substrate solution, and chromogenic substrate solution 1. Washing solution; wherein the enzyme marker is composed of alkaline phosphatase-labeled HE4 antibody and silanized horseradish peroxidase-labeled CA125 antibody;

[0064] The chemiluminescence substrate liquid is the Tris buffer solution of 0.2mol / L pH 9.7 for the chemiluminescence substrate A solution, also includes the sodium chloride (NaCl) of 2.7mol / L, the CTAC (hexadecyl chloride) of 0.08mmol / L Trimethylammonium chloride) and 2.6mmol / L of AMPPD (3-(2-spiroadamantane)-4-methoxyl-4-(3-phosphoryloxy)-phenyl-1,2-dioxo Cyclohexane disodium salt), does not contain sodium azide;

[0065] Chromogenic substrate liquid is the 0.33mol / L sodium acetate of pH 6.0±0.5 and the citric acid buffer solution ...

Embodiment 2

[0104] In this example, the kit provided in Example 1 is applied to the simultaneous detection of HE4 and CA125 in the sample, and the specific detection method is:

[0105] Ⅰ: Take the sample to be tested in the reaction cup, add the magnetic particles jointly labeled with HE4 antibody and CA125 antibody, then add the enzyme label, mix well, and incubate at 37°C for 16 minutes; wash and separate the magnetic particles; put them into the magnetic particles Add chemiluminescent substrate A solution, react and measure the light intensity; then add 100 μL of chromogenic substrate B solution to the magnetic particles, react for 5 minutes and measure the absorbance at 620 nm;

[0106]Ⅱ: Take the calibrator with known concentration and measure the light intensity and absorbance according to the method of step Ⅰ, draw the standard working curve of light intensity and HE4 concentration, and obtain the corresponding relationship between light intensity and HE4 concentration; draw the st...

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Abstract

The invention relates to technical field of immunodetection, particularly relates to a kit for joint detection of ovarian cancer tumor markers HE4 and CA125 as well as a preparation method and application thereof. According to the kit provided by the invention, silanization treatment is performed on horse radish peroxidase, so that the possibility that the horse radish peroxidase becomes a phosphoric acid receptor in a process of catalyzing a chemiluminescent substrate to emit light by a alkaline phosphatase is blocked, the problem that the existence of horse radish peroxidase disturbs the cataluminescence of the alkaline phosphatase is solved, so that the size of the light intensity caused by the alkaline phosphatase on magnetic particles is detected by chemiluminescent substrate liquid first, the size of chromogenic absorbancy caused by the horse radish peroxidase is detected by chromogenic substrate liquid and the simultaneous detection on HE4 and CA125 of a sample is realized. Thekit provided by the invention has the advantages that the detection process is simple, the reaction time is short, the reagent dose is less, the cost is reduced, the sensitivity is high, the repeatability and stability of the reagent are good.

Description

technical field [0001] The invention relates to the technical field of immune detection, in particular to a kit for combined detection of ovarian cancer tumor markers HE4 and CA125, a preparation method and application thereof. Background technique [0002] Ovarian cancer is one of the malignant tumors of the gynecological reproductive system. Its incidence ranks third in gynecological tumors after cervical cancer and uterine body cancer, but its fatality rate ranks first in gynecological diseases. The main reason is that the ovary is deeply hidden in the bone cavity, the early symptoms of ovarian cancer are not obvious, and there is no effective early diagnosis method in clinical practice. Many patients are already in the advanced stage of cancer when they are diagnosed. The five-year survival rate of early stage patients is more than 90%, while that of advanced stage patients is less than 30%. Therefore, how to diagnose ovarian cancer as early as possible is extremely cri...

Claims

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Application Information

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IPC IPC(8): G01N33/574G01N33/543G01N33/535G01N33/577
CPCG01N33/535G01N33/54326G01N33/57449G01N33/57484G01N33/577G01N2446/90G01N2800/36
Inventor 刘承举刘向祎路晟
Owner 北京惠中医疗器械有限公司
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