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Kit for detecting hepatocellular carcinoma (HCC)

A hepatocellular carcinoma and reagent kit technology, applied in the field of medical testing, can solve the problems of unsatisfactory treatment effect, limited clinical application, and low sensitivity of blood detection method, and achieve low cost, less blood consumption, and simple operation Effect

Active Publication Date: 2018-12-07
WUHAN UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Moreover, the sensitivity of the blood detection method (AFP method) is very low, which limits its clinical application.
When patients are diagnosed, they are often in the middle and late stages, and the treatment effect at this time will not be very satisfactory.

Method used

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  • Kit for detecting hepatocellular carcinoma (HCC)
  • Kit for detecting hepatocellular carcinoma (HCC)
  • Kit for detecting hepatocellular carcinoma (HCC)

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] 1. Extraction of ctDNA

[0022] Blood samples were collected in StreckBCT tubes, stored at 4 °C, and ctDNA was extracted within 24 h of collection. Using the Circulating Nucleic Acid kit of QIAamp Company, the ctDNA in plasma was extracted according to the operation steps in the manual.

[0023] 2. Detection of ctDNA length

[0024] The length of the extracted ctDNA was checked by agarose gel electrophoresis. The specific steps are:

[0025] 1) Add 1 g of agarose to 100 mL of 1×TAE running buffer and shake well. Heat in the microwave until the agarose is completely dissolved. Cool to 60°C, add 100 μL of 0.5 mg / mL EB, and shake well.

[0026] 2) Seal both ends of the glue making plate with tape, insert a comb, pour the dissolved agarose, and cool at room temperature to solidify.

[0027] 3) After fully solidified, tear off the tapes at both ends, put the gel into the electrophoresis tank, add 1×TAE electrophoresis buffer until the liquid surface covers the gel by 1-...

Embodiment 2

[0048] Double-blind testing was performed as in Example 1.

[0049] Subjects: A total of 202 subjects were enrolled from September 2017 to April 2018. The enrolled patients met the diagnosis of primary liver cancer in the "China Common Malignant Tumor Diagnosis and Treatment Standards"; the age range was 18-70 years old (including 18 years old, 70 years old); there is a clear measurable tumor; no tumor therapy such as radiotherapy and chemotherapy and immunotherapy are used in the first month of enrollment; the quality of life is greater than 60 points; the expected survival period of the subjects is greater than 3 months; understood and signed the informed consent.

[0050] According to the "Guiding Principles of Clinical Research on New Chinese Medicines" according to TCM syndrome differentiation, primary liver cancer belongs to deficiency of both spleen and kidney, and internal resistance of blood stasis and toxin. Symptoms include abdominal distention and fullness, lack o...

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Abstract

The invention discloses a kit for detecting hepatocellular carcinoma (HCC) and belongs to the technical field of medical examination. The kit comprises five pairs of primers for detecting five methylation specific sites of a genome, and the sequences of the primers are represented by SEQ ID No.1-10. Genome DNA of the blood of a person to be detected is extracted; after bisulfate processing, 5 pairs of primers are subjected to PCR amplification, the amplification products are connected to a clone carrier, cloning and sequencing are performed; according to the sequencing results, the number of methylation sites is determined; then the ctDNA of the blood of the person is extracted, and the length of the ctDNA is measured. If the number of methylation sites is 3 or more, the person has HCC; and if the number of methylation sites is 1 or 2 and the length of ctDNA is less than 245 bp, the person has HCC. The kit has the advantages that the amount of used blood is little, the operation is convenient, the cost is low, and the accuracy is high; and can be applied to early diagnosis and mass screening of HCC.

Description

technical field [0001] The invention relates to the technical field of medical testing, in particular to a kit for detecting hepatocellular carcinoma. Background technique [0002] With the development of medical technology, cancer detection technology has become more and more mature, but there are few reports on early detection methods of cancer. At present, hepatocellular carcinoma (HCC) is the most common primary liver cancer in adults and one of the main causes of cancer death worldwide. However, HCC is relatively insidious, and the current detection methods for HCC mainly rely on imaging and non-specific methods. A blood test for the tumor marker alpha-fetoprotein (AFP), which is usually elevated only when the disease is in the middle and late stages. Moreover, the sensitivity of blood detection method (AFP method) is very low, so its clinical application is very limited. Patients are often diagnosed in the middle and late stages, and the treatment effect at this time...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12N15/11
CPCC12Q1/6886C12Q2600/154
Inventor 廖兴华戴周彤项园张慧敏黄凤张子健李含含姚奥范丽娟李佳蓬杜庆李慧张同存
Owner WUHAN UNIV OF SCI & TECH
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