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Primer probe composition, kit and method for early screening of liver cancer

A primer-probe and composition technology, which can be used in biochemical equipment and methods, recombinant DNA technology, and microbial assay/inspection, etc., can solve problems such as prevention and early screening of unwell liver cancer, achieve high sensitivity and specificity, improve Good accuracy and sensitivity

Pending Publication Date: 2022-01-18
苏州美汇医药技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although real-time fluorescence quantitative PCR has been applied to the screening of liver cancer at present, the accuracy and sensitivity of its detection need to be further improved, and it is not suitable for the prevention and early screening of liver cancer.

Method used

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  • Primer probe composition, kit and method for early screening of liver cancer
  • Primer probe composition, kit and method for early screening of liver cancer
  • Primer probe composition, kit and method for early screening of liver cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Embodiment 1: Kit

[0044] The kit is an early screening and prognosis monitoring kit for liver cancer, and a kit for simultaneously detecting the methylation of two genes in blood cells. The two genes and the internal reference ACTB gene are combined to form a triple nucleic acid reaction solution. The kit includes nucleic acid reaction solution A and nucleic acid reaction solution B, wherein nucleic acid reaction solution A includes a primer-probe composition, and nucleic acid reaction solution B is purchased qPCR PreMix (Probe). See Table 1 for specific information on the primers and probes of the nucleic acid reaction solution A.

[0045] Table 1 The sequence of the primer probe composition

[0046]

[0047] Among them, the 5' end of probe P1 is labeled with a FAM fluorescent reporter group, and the 3' end is labeled with a BHQ-1 fluorescent quencher group; the 5' end of probe P2 is labeled with a VIC fluorescent reporter group, and the 3' end is labeled with T...

Embodiment 2

[0062] Embodiment 2: Sensitivity experiment

[0063] The sensitivity of the kit of Example 1 was tested. Among them, three gradients of 50%, 5% and 1% were set for the methylation rate of CDKN2A gene and GSTP1 gene. For test results, see figure 1 and image 3 As shown, the results show that the kit has good sensitivity and can detect 1% of the liver cancer gene methylation rate in the sample.

Embodiment 3

[0064] Embodiment 3: specificity experiment

[0065] The kit of Example 1 was used to detect 23 clinical samples, including 5 samples of fatty liver, 7 samples of hepatitis and 11 samples of liver cancer. The kit was used to detect each sample separately, and the sample volume of each sample DNA was 14 μL. The reaction procedure is as in Table 3 above.

[0066] The information of 27 clinical samples is shown in Table 4.

[0067] Table 4 Clinical sample information

[0068]

[0069]

[0070] Calculate the sensitivity and specificity of the kit according to Table 4 as follows:

[0071] Sensitivity: number of detected liver cancer / total number of liver cancer×100%=(9 / 11)×100%=81.82%.

[0072] Specificity: Number of detected non-bladder cancer / total number of non-bladder cancer×100%=(12 / 12)×100%=100%.

[0073] The kit and detection method have good sensitivity and specificity.

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Abstract

The invention discloses a primer probe composition, a kit and a method for early screening of the liver cancer. The primer probe composition comprises: a forward primer F1, the nucleotide sequence of which is as shown in SEQ ID NO.1; a reverse primer R1, the nucleotide sequence of which is as shown in SEQ ID NO. 2; a probe P1, the nucleotide sequence of which is as shown in SEQ ID NO.3; a forward primer F2, the nucleotide sequence of which is as shown in SEQ ID NO. 4; a reverse primer R2, the nucleotide sequence of which is as shown in SEQ ID NO.5; and a probe P2, the nucleotide sequence of which is as shown in SEQ ID NO. 6. The primer probe composition is high in specificity, high in sensitivity, relatively reliable and suitable for prevention, early screening and prognosis monitoring of the liver cancer.

Description

technical field [0001] The invention belongs to the field of liver cancer detection kits, and relates to a primer probe composition, a kit and a method for early screening of liver cancer, in particular to a methylation fluorescent quantitative PCR-based early screening kit for liver cancer. Background technique [0002] Liver cancer is a highly vascularized tumor that ranks 2nd in cancer deaths. The number of deaths from liver cancer is increasing year by year. In China, the number of deaths from liver cancer is 383,000 per year, accounting for 51% of the world's liver cancer deaths. 50% of patients diagnosed with liver cancer died within 1 year, and the average survival time of less than 6% of patients was 5 years. The most common risk factors for liver cancer include smoking, alcohol consumption, aflatoxins, metabolic syndrome, and long-term chronic infection with hepatitis B virus (HBV) or hepatitis C virus (HCV). In recent years, the incidence of obesity and type 2 di...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12Q1/6851C12N15/11
CPCC12Q1/6886C12Q1/6851C12Q2600/166C12Q2600/154C12Q2531/113C12Q2523/125C12Q2545/101C12Q2563/107
Inventor 颜菁袁青赵荣荣周志鹏
Owner 苏州美汇医药技术有限公司
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