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Combined detection serum marker for early screening and diagnosis of liver cancer, kit and detection method

A technology of combined detection of serum markers, applied in disease diagnosis, biological testing, measuring devices, etc., can solve problems such as unsatisfactory results, and achieve low cost, simple operation, and high sensitivity

Active Publication Date: 2019-09-27
ZHENGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the next work, the researchers tried to find more sensitive and specific anti-TAA autoantibodies, but the results were not satisfactory

Method used

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  • Combined detection serum marker for early screening and diagnosis of liver cancer, kit and detection method
  • Combined detection serum marker for early screening and diagnosis of liver cancer, kit and detection method
  • Combined detection serum marker for early screening and diagnosis of liver cancer, kit and detection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] The combined detection serum markers used in this example for the early screening and diagnosis of liver cancer consist of proteins encoded by six genes including PTEN, PTCH1, IDH1, SRSF2, MSH2 and NPM1. Wherein the protein encoded by the PTEN gene has the amino acid sequence shown in SEQ ID NO.2, the protein encoded by the PTCH1 gene has the amino acid sequence shown in SEQ ID NO.3, and the protein encoded by the IDH1 gene has the amino acid sequence shown in SEQ ID NO.4 The amino acid sequence of the protein encoded by the SRSF2 gene has the amino acid sequence shown in SEQ ID NO.5, the protein encoded by the MSH2 gene has the amino acid sequence shown in SEQ ID NO.6, and the protein encoded by the NPM1 gene has the amino acid sequence shown in SEQ ID NO. . The amino acid sequence shown in 7.

Embodiment 2

[0048] The kit for the early screening and diagnosis of liver cancer in this example includes the combined detection serum markers of the above-mentioned Example 1, and is coated on a polyvinyl chloride concave-well plate. In addition, the kit also includes a certain amount of positive control serum, negative control serum, blocking solution, sample diluent, secondary antibody, secondary antibody diluent, washing solution, color developing solution and stop solution, and the positive control serum is ELISA The OD value of the experiment is higher and the corresponding antibody-positive serum is verified by Western Blot experiment. The negative control serum is the serum with the OD value of the ELISA experiment in the normal control population near the average value and negative by Western Blot experiment. The second antibody is HRP. Labeled mouse anti-human IgG.

Embodiment 3

[0050] The detection method of this embodiment uses the combined detection of serum markers in the above-mentioned embodiment 1, and the specific steps are as follows:

[0051] 1) Coating: The combined detection serum markers were coated respectively (see Table 1 below for the coating concentration), 100 μL / well, overnight at 4°C.

[0052] 2) Blocking: 2% BSA in PBST (PBS, Tween20) solution, 200 μL / well, overnight at 4°C.

[0053] 3) Washing: wash 3 times with 350 μL / well PBST.

[0054] 4) Primary antibody incubation: the serum to be tested was diluted 1:100 with PBST containing 1% BSA, 100 μL / well, and placed in a semi-water bath at 37°C for 1 hour.

[0055] 5) Washing: wash 5 times with 350 μL / well PBST.

[0056] 6) Secondary antibody incubation: HRP-labeled mouse anti-human IgG was diluted 1:10000 with PBST containing 1% BSA, 100 μL / well, in a semi-water bath at 37°C for 1 hour.

[0057] 7) Washing: wash 5 times with 350 μL / well PBST.

[0058] 8) Color development: TMB ...

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Abstract

The invention relates to a combined detection serum marker for the early screening and diagnosis of liver cancer, a kit containing the combined detection serum marker and a detection method and belongs to the technical field of biomedicine. According to the combined detection serum marker for the early screening and diagnosis of the liver cancer of the invention, a human protein chip coded by 138 cancer-driven genes is customized on the basis of a role played by cancer-driven genes in tumor generation and development; an early-stage detection serum marker for liver cancer is preliminarily screened out through the protein chip; and finally a set of liver cancer combined detection serum marker which can be used for the early screening and diagnosis of liver cancer is screened out through the verification of an ELISA experiment; and the liver cancer combined detection serum marker includes a PTEN encoded protein, a PTCH1 encoded protein, an IDH1 coded protein, an SRSF2 coded protein, an MSH2 coded protein and an NPM1 coded protein. The combined detection serum marker can assist in the clinical diagnosis of liver cancer, and has the advantages of high sensitivity, strong specificity, low cost and the like.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a combined detection serum marker for early screening and diagnosis of liver cancer, a kit containing the combined detection serum marker and a detection method. Background technique [0002] Primary liver cancer is one of the most common malignant tumors in the world, which seriously threatens people's life and health. Liver cancer has an insidious onset and a poor prognosis. Surgical resection and liver transplantation are considered to be the means of radical treatment of liver cancer, but most patients have already entered the advanced stage of liver cancer when they are first diagnosed, thus losing the opportunity for treatment. The main reason is that patients with early liver cancer usually have no Obvious symptoms, or small size, are difficult to detect by imaging examinations. As the main serum marker, alpha-fetoprotein (AFP) has the disadvantage of low ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/574
CPCG01N33/6854G01N33/57438G01N2800/085
Inventor 张建营叶华代丽萍王晓王鹏史健翔王科妍杨倩
Owner ZHENGZHOU UNIV
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