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A kit for early screening of liver cancer and its application method

A kit and screening technology, applied in the field of molecular biology, can solve the problems of unsatisfactory diagnostic sensitivity and specificity, and achieve the effect of reducing diagnostic costs, high sensitivity, and rapid diagnosis

Active Publication Date: 2021-10-26
ANHUI DAJIAN MEDICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the diagnosis of primary liver cancer, the most widely used clinical tumor marker is alpha-fetoprotein (AFP), but its diagnostic sensitivity and specificity are not satisfactory, and the sensitivity and specificity of diagnosing liver cancer are 41%~ 65% and 80% to 94%

Method used

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  • A kit for early screening of liver cancer and its application method
  • A kit for early screening of liver cancer and its application method
  • A kit for early screening of liver cancer and its application method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Embodiment 1SCAND3 and its primers and probes

[0032] The present invention finds for the first time that SCAND3 methylation can be used for early screening and diagnosis of liver cancer.

[0033] The primer sequences used to detect SCAND3 methylation are as follows:

[0034] The forward primer sequence is: 5'-GTTATAAATTGAGCGGTAAGATATTTGC-3', as shown in the sequence of SEQ ID NO: 1;

[0035] The reverse primer sequence is: 5'-CCTCGCCCAAACTACTCCG-3', as shown in SEQ ID NO: 2 sequence;,

[0036] The fluorescent probe sequence is: 5'-AATCGAGGTTATGACGGATA-3', as shown in the sequence of SEQ ID NO: 3; the 5' end of the fluorescent probe is marked with FAM (fluorescent reporter group), and the 3' end is marked with MGB (fluorescent quencher group). group).

[0037] Other primers and probes used in the screening process of the present invention are as follows:

[0038] The first set of primers and probes:

[0039] The forward primer sequence is shown in SEQ ID NO.4 (5'-...

Embodiment 2

[0042] Example 2 A kit for early screening of liver cancer

[0043] The kit includes a forward primer whose sequence is shown in SEQ ID NO.1, a reverse primer whose sequence is SEQ ID NO.2 and a fluorescent probe whose sequence is SEQ ID NO.3; the 5' end of the probe Labeled FAM, 3′ end labeled MGB; In addition, it also includes forward primers and probes for detecting GAPDH, PCR Master Mix, lysate, proteinase K, washing solution, bisulfite solution and ddH 2 O: Forward primers and probes for detecting GAPDH can be primers and probes known in the art.

Embodiment 3

[0044] The using method of kit described in embodiment 3

[0045] The using method of the kit described in embodiment 2 is as follows:

[0046] (1) Extraction of genomic DNA from peripheral blood cells: take peripheral blood cells, use lysate, proteinase K, rinse solution and ddH 2 O is extracted to obtain a genomic DNA solution;

[0047] The specific steps are:

[0048] Aspirate 200 μL of peripheral blood sample, put it into a 1.5mL centrifuge tube, add 1mL of lysate 1, then continue to vortex and oscillate for 1min, mix thoroughly, and then place the 1.5mL centrifuge tube in a 70°C water bath for 5min.

[0049] Take the 1.5mL centrifuge tube out of the water bath, centrifuge at 12,000rpm for 5min, and slowly pipette 300μL of the supernatant into a new 1.5mL centrifuge tube.

[0050] Add 200 μL Lysis Solution 2 and 20 μL Proteinase K to a new 1.5 mL centrifuge tube in sequence, and vortex to mix.

[0051] Place a new 1.5mL centrifuge tube in a 70°C water bath for 10min.

...

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Abstract

The invention belongs to the field of molecular biology, and in particular relates to a kit for early screening of liver cancer and a use method thereof. The present invention finds for the first time that SCAND3 can be used as a methylation marker for detecting liver cancer. The primer sequence for detecting SCAND3 methylation is as follows: the forward primer sequence is shown in SEQ ID NO.1, the reverse primer sequence is shown in SEQ ID NO.2; the fluorescent probe sequence is SEQ ID NO.3, and the probe The 5' end of the needle is labeled FAM and the 3' end is labeled MGB. The present invention uses SCAND3 as a methylation marker for early screening of liver cancer for diagnosis of liver cancer, which has the advantages of high sensitivity and strong specificity, especially in patients with AFP<400 μg / L, the incidence of SCAND3 methylation As high as 60%. It can be seen that the present invention uses SCAND3 as a methylation marker and is suitable for the diagnosis of liver cancer, especially for the diagnosis of early liver cancer.

Description

technical field [0001] The invention belongs to the field of molecular biology, and in particular relates to a kit for early screening of liver cancer and a use method thereof. Background technique [0002] Hepatocellular carcinoma (HCC) is one of the common malignant tumors that threaten human life, and most patients with primary liver cancer have no obvious early symptoms, and most patients have reached the middle and late stages when they are diagnosed, so early diagnosis and curative treatment can be achieved. Resection is the patient's only hope for long-term survival. Due to the lack of specific tumor markers, most patients are diagnosed at an advanced stage and lose the chance of radical resection. [0003] At present, the diagnostic methods of liver cancer include imaging examination, pathological examination and laboratory examination. However, the imaging manifestations of early liver cancer are not typical, and some examinations are radioactive and expensive. L...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6886C12N15/11
CPCC12Q1/6886C12Q2600/154
Inventor 邵琦
Owner ANHUI DAJIAN MEDICAL TECH CO LTD
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