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Composition for detecting liver cancer and application thereof

A composition and liver cancer technology, applied in the biological field, can solve the problems of unsatisfactory specificity and sensitivity, high equipment cost, and increased AFP concentration, and achieve the effect of an accurate in vitro detection method

Inactive Publication Date: 2016-12-14
BIOCHAIN BEIJING SCI & TECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The current technology used for liver cancer detection has very limited application in the detection and screening of early liver cancer: 1) Imaging detection technologies (B-ultrasound, CT, MRI, etc.) have high equipment costs, strong operational technical requirements, and high detection costs; 2 ) tissue biopsy is highly invasive and not suitable for early cancer screening
Moreover, such a invasive detection method brings great physical pain to the patient, so it is not easy for the patient to accept
[0006] Serum tumor markers (CEA, CA125, and CA199, etc.) are methods developed in recent years for the detection of diseases. Although they have a certain role as a non-invasive detection method for liver cancer, they still cannot fully meet the needs of clinical detection due to their low sensitivity. need
[0007] The serum marker that is widely used in the detection of liver cancer is alpha-fetoprotein (AFP). As a commonly used tumor molecular marker, it has good detection specificity, but only about 60% of patients with primary liver cancer have AFP concentration. In other words, about 40% of liver cancer patients are AFP-negative, and in new cases, the proportion of AFP-negative liver cancer continues to increase, and the specificity and sensitivity of AFP detection of liver cancer can no longer meet current needs

Method used

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  • Composition for detecting liver cancer and application thereof
  • Composition for detecting liver cancer and application thereof
  • Composition for detecting liver cancer and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0101] Example 1: Detection of methylated DNA of SEPT9 gene.

[0102] Liver cancer, liver disease and normal people were detected. For the probes and primers used by SEPT9, you can refer to the prior art, such as the probes and primers recorded in Chinese patent CN101160411A (equivalent to SEQ ID NO: 2-5 of the present invention)

[0103] This embodiment includes the following steps:

[0104] First, plasma samples from 15 patients with liver cancer, 24 patients with liver disease (12 in the compensated phase and 12 in the decompensated phase) and 10 normal individuals were obtained. All samples come from Boercheng Company. Then the genomic DNA of liver cancer and normal people is extracted, and the genomic DNA samples are pretreated so that the unmethylated cytosine base at the 5' position is converted into uracil, thymine or not used in the hybridization behavior. Another base for pyrimidine. In this example, this pretreatment was achieved by bisulfite reagent treatment...

Embodiment 2

[0108] Example 2: Detection of methylated DNA of SEPT9 gene, detection of AFP-negative liver cancer patients

[0109] First, plasma samples from 15 liver cancer patients were obtained. All samples come from Boercheng Company. Then the genomic DNA of the plasma sample of the liver cancer patient is extracted, and the genomic DNA sample is pretreated so that the unmethylated cytosine base at the 5' position is converted into uracil, thymine or not used in the hybridization behavior. Another base for pyrimidine. In this example, this pretreatment was achieved by bisulfite reagent treatment. The extraction and processing of the DNA can be carried out by any standard means in the prior art. Specifically, in this embodiment, all sample DNA extraction and bisulfite DNA modification are carried out by using Boercheng Company Extracted from the plasma processing kit. Then, the above-mentioned SEPT9 gene primer, probe combination and the internal reference gene ACTB gene primer an...

Embodiment 3

[0113] Example 3: Joint detection of various markers in patients with liver cancer

[0114] The pathogenesis, pathogenesis, and clinical manifestations of HCC are highly heterogeneous. Therefore, the effect of using a single marker for detection is not ideal. Based on this, the inventors detected liver cancer through joint detection of multiple markers.

[0115] First, plasma samples from 10 liver cancer patients were obtained. All samples come from Boercheng Company. Then, serum markers were detected on the serum samples of these 15 patients with liver cancer, three different serum markers of CEA, CA125 and AFP were detected, and SEPT9 methylation was detected at the same time. The method of SEPT9 methylation detection As shown in Example 1 and Example 2.

[0116]Extract the genomic DNA of the plasma sample of the liver cancer patient, and pretreat the genomic DNA sample so that the unmethylated cytosine base at the 5' position is converted into uracil, thymine or not u...

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Abstract

The invention provides a composition for detecting liver cancer, which comprises a nucleic acid for detecting methylation status in at least one zone of SEPT9 gene and fragments thereof and a reagent for detecting concentration of alpha fetoprotein in serum, so as to detect the liver cancer by means of the methylation status of the gene and the fragments thereof and the concentration of the alpha fetoprotein. The invention also provides a kit comprising the composition and an in-vitro detection method for the liver cancer. In the invention, the DNA in a plasma sample is analyzed by real-time PCR, so as to conveniently detect the SEPT9 gene and the fragments thereof, and according to the circulating threshold value (Ct) of the real-time PCR, the sample is conveniently and quickly determined to be positive or negative; and furthermore, the invention provides the accurate in-vitro detection method by detecting the concentration of the alpha fetoprotein in the sample.

Description

technical field [0001] The invention belongs to the field of biotechnology and relates to a composition and its use in disease detection, in particular to a composition for detecting liver cancer and its corresponding kit and use. Background technique [0002] Cancer is a disease that seriously threatens public health. The latest data on the global cancer situation (GLOBOCAN2012) released by the International Agency for Research on Cancer under the World Health Organization shows that in 2012, there were about 14.1 million new cancer cases worldwide, and the number of cancer deaths reached 8.2 million; compared with the data in 2008, New cancer cases increased by 11% in 2012, and cancer deaths increased by 8.3%. Based on available data, the agency predicts that due to global population growth and aging, there will be as many as 19.3 million new cancer cases worldwide each year by 2025. [0003] Due to the aging population and changes in people's lifestyles in recent years,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68G01N33/574
CPCC12Q1/6886C12Q2600/154G01N33/57438
Inventor 马竣周光朋韩晓亮王建铭宋乐乐徐海
Owner BIOCHAIN BEIJING SCI & TECH
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