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A kit for hepatitis B related hepatocellular carcinoma early diagnosis

An early diagnosis and correlation technology, applied in biological testing, material inspection products, measurement devices, etc., can solve problems such as loss of surgical treatment opportunities, difficulty in early diagnosis, hidden incidence of HCC, etc., to avoid operation errors and environmental impact, simplifying Operation, the effect of improving detection efficiency

Active Publication Date: 2017-02-08
惠森生物科技(上海)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The onset of HCC is hidden, and early diagnosis is difficult. Most patients are in the middle and late stages when they are clinically diagnosed, thus losing the best opportunity for surgical treatment.
Alpha fetoprotein (AFP) is currently the most commonly used diagnostic marker for HCC clinically, but its sensitivity has certain limitations, and more than 30% of HCC patients have negative AFP detection results

Method used

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  • A kit for hepatitis B related hepatocellular carcinoma early diagnosis
  • A kit for hepatitis B related hepatocellular carcinoma early diagnosis
  • A kit for hepatitis B related hepatocellular carcinoma early diagnosis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] The composition of embodiment 1 hepatitis B related HCC diagnostic kit

[0036] (1) At least one microporous plate; the microporous plate may include a Gal-3BP joint AFP quantitative detection microporous plate, which is composed of a Gal-3BP-ELISA part and an AFP-ELISA part, and its arrangement is as follows figure 2 Shown; Wherein, the microwell of Gal-3BP-ELISA part is coated with anti-Gal-3BP capture antibody, and the microwell of AFP-ELISA part is coated with anti-AFP capture antibody;

[0037] (2) Anti-Gal-3BP capture antibody, anti-Gal-3BP detection antibody, anti-AFP capture antibody and anti-AFP detection antibody;

[0038] (3) Serum sample diluent: 1mM EDTA, 0.5% Triton X-100 in PBS, pH 7.2-7.4;

[0039] (4) Standard products: Gal-3BP (50ng / bottle) and AFP (200ng / bottle);

[0040] (5) Blocking agent: 1% BSA, 0.05% NaN 3 PBS solution, pH7.2-7.4;

[0041] (6) Cleaning solution: PBS solution of 0.05% Tween 20, pH7.2-7.4;

[0042] (7) Chromogenic solution: TMB...

Embodiment 2

[0049] Example 2 Preparation and pretreatment of hepatitis B-related HCC diagnostic kit Gal-3BP-ELISA plate and AFP-ELISA plate

[0050] (1) Take a blank 96-well microwell plate, with 48 wells in the upper and lower parts, and use it as the Gal-3BP-ELISA plate and the AFP-ELISA plate respectively (such as figure 2 );

[0051] (2) The anti-Gal-3BP capture antibody (2 μg / mL) and anti-AFP capture antibody (2 μg / mL) were added to the Gal-3BP-ELISA plate and the AFP-ELISA plate respectively, 0.1 mL per well, and incubated at 37 ° C for 2 hours or overnight at 4°C.

[0052] (3) Wash the above-mentioned microwell plate with cleaning solution, 300 μL / well, soaking time is 2 minutes / time, wash four times in total and pat dry the remaining cleaning solution in the well.

[0053] (4) Seal the above-mentioned microwell plate with a sealing agent, 200 μL / well, and seal at room temperature for 1 hour; place it in a drying room at 25-35°C and humidity <40% for 20-24 hours to obtain Gal-3B...

Embodiment 3

[0054] The drawing of embodiment 3 standard curve

[0055] Dissolve and dilute the Gal-3BP and AFP standard substance with PBS, so that the concentrations of the Gal-3BP standard substance are respectively: 25ng / mL, 12.5ng / mL, 6.25ng / mL, 3.13ng / mL, 1.56ng / mL, 0.781 ng / mL, 0.391ng / mL, 0ng / mL, so that the concentrations of AFP standards are: 20ng / mL, 10ng / mL, 5ng / mL, 2.5ng / mL, 1.25ng / mL, 0.625ng / mL, 0.313 ng / mL, 0 ng / mL.

[0056] The detection steps are as follows:

[0057] (1) Numbering: Number the corresponding microwells of the standard in sequence.

[0058] (2) Adding samples: Add 50 μL / well of the standard solution to the corresponding wells, shake gently to mix.

[0059] (3) Incubation: seal the plate with a plate sealing film and incubate at room temperature for 2 hours.

[0060] (4) Washing: spin dry the liquid in the wells, wash with cleaning solution, 300 μL / well, soak for 2 minutes / time, wash four times in total and pat dry the remaining cleaning solution in the w...

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Abstract

The invention discloses an enzyme-linked immunosorbent assay kit which is rapid, simple, convenient, and high in diagnosis sensitivity and accuracy for hepatitis B related HCC early diagnosis. The kit includes an anti-Gal-3BP capture antibody, an anti-Gal-3BP detection antibody, an anti-AFP capture antibody and an anti-AFP detection antibody. The kit can achieve comprehensive consideration of Gal-3BP and AFP expression levels in serum of a tested person, and increases the diagnosis sensitivity and accuracy, and therefore a patient can know the disease risk in an early stage of the disease, and corresponding treatment measures can be taken in time. The kit has a wide application prospect.

Description

technical field [0001] The invention belongs to the field of molecular diagnosis and relates to a kit for early diagnosis of hepatitis B-related hepatocellular carcinoma. Background technique [0002] As one of the malignant tumors with high morbidity and mortality worldwide, hepatocellular carcinoma (HCC) has the characteristics of high malignancy, rapid progression, poor prognosis and strong invasiveness. About 50% of the new and dead HCC cases in the world occur in my country every year. Since the first report in 1975 that hepatitis B virus (HBV) infection was closely related to the occurrence of HCC, successive studies have confirmed that the global distribution of hepatitis B surface antigen (HBsAg) positive population is closely related to the occurrence of HCC. . my country is a high-incidence area of ​​chronic hepatitis B, and more than half of the occurrence and death of HCC are attributed to HBV infection. [0003] The onset of HCC is occult, and early diagnosis...

Claims

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Application Information

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IPC IPC(8): G01N33/574G01N33/68
CPCG01N33/574G01N33/57438G01N33/68
Inventor 郭坤刘天华张舒刘银坤
Owner 惠森生物科技(上海)有限公司
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