Composition for detecting liver cancer as well as kit and application thereof

A detection kit and kit technology, applied in the field of liver cancer detection compositions, can solve the problems of only targeting a single gene, limited diagnostic effect, and unsatisfactory detection accuracy, achieving high accuracy, high sensitivity, and early detection. Effectiveness of screening and diagnosis

Pending Publication Date: 2021-12-10
广州达健生物科技有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are few methods for detecting DNA methylation in liver cancer based on methylation fluorescence quantitative method. At the same time, relevant detection is often only for a single gene, and the detection accuracy is often not as ideal, and the diagnostic effect is limited.

Method used

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  • Composition for detecting liver cancer as well as kit and application thereof
  • Composition for detecting liver cancer as well as kit and application thereof
  • Composition for detecting liver cancer as well as kit and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1: Sample DNA Extraction and Bisulfite Conversion

[0042] 1. Serum sample processing and DNA extraction

[0043] 1) Acquisition of samples to be tested: select serum samples from clinical patients: take the venous blood of the subject, draw 2mL with a sterile injection needle, collect it in a sterile collection tube, and place it at room temperature for 30 minutes. Use a horizontal centrifuge, centrifuge at 3000rpm for 5min, absorb the serum and transfer it to a 1.5mL centrifuge tube for later use.

[0044] 2) DNA extraction:

[0045] The nucleic acid extraction kit of Guangzhou Dajian Biotechnology Co., Ltd. was used for extraction, and the operation was as follows:

[0046] (a) Take a 1.5mLEP tube, add 200μL of the sample to be tested and 20μL of pancreatic lipase in turn, vortex, mix well, and place at room temperature for 5min;

[0047] (b) Add 20 μL of proteinase K and 360 μL of lysate to the EP tube, vortex, mix thoroughly, centrifuge briefly, and bath...

Embodiment 2

[0066] Example 2: Screening of candidate genes for hypermethylation in liver cancer tissue and specific primers and probes

[0067] 1. Screening of serum hypermethylated candidate genes in patients with liver cancer

[0068] Through the TCGA database (http: / / cancergenome.nih.gov / ), the methylation microarray data related to liver cancer were obtained for analysis, and 9 candidate gene sites with high methylation in liver cancer were obtained: SGIP1, SCAND3, MYO1G, CDKN2A, Slit2, DAPK, PSD4, KLF3, ATXN1. Finally, we screened out SGIP1, SCAND3, MYO1G.

[0069] 2. Screening of specific primers and probes for methylation detection of liver cancer

[0070] 1) Design and screening of specific primers and probes:

[0071] According to the nucleic acid sequences of SGIP1, SCAND3, and MYO1G mentioned above, the methylation primers and probes were designed on the Methyl primer Express v1.0 software. After repeated design and deliberation by the applicant, the fluorescent quantitative...

Embodiment 3

[0087] Example 3: Clinical sample detection and verification kit effect

[0088] 1. Blood sample liver cancer gene methylation detection results:

[0089] In order to evaluate that the probes and primers provided by the present invention can be used to detect the methylation of liver cancer SGIP1, SCAND3, and MYO1G genes in samples by fluorescent PCR, the DNA templates derived from the same sample were divided into 12 parts and completed under different detection systems of T1-T12 The detection system of its fluorescent PCR is shown in the following table:

[0090] Numbering Primer Probe Set T1 SGIP1-1 T2 SGIP1-3 T3 SCAND3-2 T4 SCAND3-3 T5 MYO1G-2 T6 MYO1G-3 T7 SGIP1-1, SCAND3-2, MYO1G-3 T8 SGIP1-1, SCAND3-3, MYO1G-2 T9 SGIP1-1, SCAND3-3, MYO1G-3 T10 SGIP1-3, SCAND3-2, MYO1G-2 T11 SGIP1-3, SCAND3-2, MYO1G-3 T12 SGIP1-3, SCAND3-3, MYO1G-3

[0091] In the T1-T12 fluorescent PCR detection sys...

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Abstract

The invention provides a composition for detecting liver cancer as well as a kit and application thereof. The liver cancer gene methylation detection kit provided by the invention takes an important biological index DNA methylation abnormality of cancer diagnosis, early screening and prognosis as a detection object, can realize non-invasive detection by detecting the gene methylation state of peripheral blood, and simultaneously adopts a fluorescent quantitative PCR technology, methylation chip data related to liver cancer is obtained through TCGA data for analysis, methylation detection sites of three genes such as SGIP1, SCAND3 and MYO1G are screened out, and the liver cancer detection kit with higher sensitivity and better specificity is obtained by establishing liver cancer-based methylation detection based on fluorescent quantitative PCR, so that early screening and diagnosis of liver cancer are realized, and early diagnosis and early treatment of liver cancer are facilitated.

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 application. Background technique [0002] Liver cancer is one of the most common malignant tumors in my country, and its mortality rate ranks second among malignant tumors. Usually occurs in middle-aged men. Liver cancer usually has no symptoms or atypical symptoms in the early stage. There may be non-specific gastrointestinal symptoms such as loss of appetite, abdominal distension, and vomiting. When patients feel obvious discomfort or clinical symptoms are very obvious, the disease mostly enters In the middle and advanced stages, the treatment of advanced liver cancer is not ideal, and the survival time is often only half a year to one and a half years. Therefore, the establishment of early warning and early screening methods for liver cancer is ver...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886C12Q1/6858C12N15/11
CPCC12Q1/6886C12Q1/6858C12Q2600/154C12Q2600/118C12Q2600/166C12Q2531/113C12Q2563/107C12Q2523/125C12Q2545/101C12Q2545/113
Inventor 邵琦
Owner 广州达健生物科技有限公司
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