Multi-element generic composition and use thereof

A technology of composition and application, which is applied in the field of kits for detecting abnormal cell proliferation in individuals, can solve problems such as poor effect and side effects, achieve the effect of improving sensitivity and specificity, and ensuring correctness and reliability

Active Publication Date: 2015-07-01
BIOCHAIN BEIJING SCI & TECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although chemotherapy has become one of the indispensable methods for the treatment of gastric cancer, it is often accompanied by side effects of different severity, and the effect is not good

Method used

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  • Multi-element generic composition and use thereof
  • Multi-element generic composition and use thereof
  • Multi-element generic composition and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0195] Example 1: Primer and probe screening

[0196] Because according to some specific embodiments, because the kit for measuring the methylation status of at least one CpG dinucleotide of the gene DNA sequence of Septin9 was purchased from Epigenomics (Germany), it can be directly carried out according to the instructions of the kit. Get primers and probes for genetic assays targeting Septin9. For the RNF180 gene, many combinations of probes and primers can be designed, and the performance of each combination of probes and primers may be different. Therefore, in the following examples, the probes and primers were screened.

[0197] In this example, the primers and probes of RNF180 were first screened with artificial methylated templates and unmethylated templates. This embodiment includes the following steps:

[0198] First, design various primers and probes of RNF180, as long as they can be equal to, complementary to or hybridize under moderate stringency or stringent...

Embodiment 2

[0227] Example 2: In vitro test of RNF180 and Septin9 on cancer and healthy human leukocyte DNA.

[0228] Samples from 4 gastric cancers (S1-S4), 2 intestinal cancers (C1-C2), 2 lung cancers (L1-L2), 1 blood cancer (Jurkat) and 7 normal people (WBC1-7) were obtained. Genomic DNA was extracted from 4 cases of gastric cancer, 2 cases of intestinal cancer, 2 cases of lung cancer, 1 case of blood cancer and 7 cases of normal people. All cancer samples were from Boercheng Company. Human normal samples were from BioReclamationIVT Company. The DNA extraction can be performed by any standard means in the prior art. Specifically, in this example, DNA from all human samples is extracted by using EPi proColon Plasma Quick Kit from Epigenomics.

[0229] Then, the genomic DNA sample is pretreated so that the unmethylated cytosine base at the 5' position is converted to uracil, thymine, or another base not available for cytosine in terms of hybridization behavior. In this example, this...

Embodiment 3

[0240] Example 3: Preliminary clinical results of multiple detection of methylated DNA of Septin9 and RNF180 in plasma of gastric cancer patients

[0241] Samples from 10 patients with gastric cancer and 11 healthy people were obtained. Genomic DNA was extracted from 10 patients with gastric cancer and 11 healthy individuals. All cancer samples were from Boercheng Company. Human normal samples were from BioReclamationIVT Company. The DNA extraction can be performed by any standard means in the prior art. Specifically, in this example, all sample DNA is extracted by using EPi proColon Plasma Quick Kit from Epigenomics.

[0242] Then, the genomic DNA sample is pretreated so that the unmethylated cytosine base at the 5' position is converted to uracil, thymine, or another base not available for cytosine in terms of hybridization behavior. In this example, this pretreatment was achieved by bisulfite reagent treatment. Modification of bisulfite DNA was performed using the EPi...

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Abstract

The invention provides use of a composition for preparing a kit for detecting cell proliferative abnormalities of an individual. The composition comprises a nucleic acid for detecting the methylation level of Septin9 and RNF180 genes as well as at least one target region in the fragments of the genes. Therefore, existence of cell proliferative abnormalities is indicated by virtue of integrating the methylation detection results of Septin9 and RNF180.

Description

technical field [0001] The present invention relates to gene detection, in particular to the use of a composition in the preparation of a kit for detecting abnormal cell proliferation in individuals. Background technique [0002] Diseases caused by abnormal cell proliferation, such as cancer, are a major public health problem, accounting for approximately 25% of all deaths in the United States. Among them, gastric cancer is the fourth most commonly diagnosed cancer in the world, and ranks the second highest in the death rate among all cancers, and is regarded as one of the biggest health nemesis in the world. Especially in many Asian countries, including China, gastric cancer ranks first among various malignant tumors, and its morbidity and mortality are very high. [0003] Moreover, gastric cancer often does not have any special symptoms in the early stage, leading to the fact that most patients diagnosed with gastric cancer are already at an advanced stage when they are d...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12N15/11C12Q1/68
Inventor 王建铭吴开春韩晓亮聂勇战
Owner BIOCHAIN BEIJING SCI & TECH
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