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Kit for early screening of liver cancer and using method of kit

A kit and liver cancer technology, applied in the field of molecular biology, can solve the problems of unsatisfactory diagnostic sensitivity and specificity, and achieve the effects of reducing diagnostic costs, reducing pain, and rapid diagnosis

Active Publication Date: 2018-11-06
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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  • Kit for early screening of liver cancer and using method of kit
  • Kit for early screening of liver cancer and using method of kit
  • Kit for early screening of liver cancer and using method of kit

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 particularly relates to a kit for early screening of liver cancer and a using method of the kit. The invention firstly discovers that SCAND3can serve as a methylation marker for detecting the liver cancer. Sequences of primers for detecting SCAND3 methylation are as follows: the sequence of a forward primer is shown as SEQ ID NO. 1; thesequence of a reverse primer is shown as SEQ ID NO. 2; the sequence of a fluorescence probe is SEQ ID NO. 3; the 5' terminal of the probe is labeled with FAM, and the 3' terminal of the probe is labeled with MGB. The SCAND3 is the methylation marker for early screening of the liver cancer, is used for liver cancer diagnosis, and has the advantages of high sensitivity and strong specificity, and particularly in a patient having the AFP (Alpha-Fetoprotein) of less than 400 ug / L, the SCAND3 methylation incidence reaches 60%. Therefore, the kit disclosed by the invention takes the SCAND3 as the methylation marker, is applicable to diagnosis of the liver cancer, and is particularly applicable to 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 Applications(China)
IPC IPC(8): C12Q1/6886C12N15/11
CPCC12Q1/6886C12Q2600/154
Inventor 邵琦
Owner ANHUI DAJIAN MEDICAL TECH CO LTD
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